ORIGINAL RESEARCH ARTICLE

published: 26 November 2014

NEUROANATOMY doi: 10.3389/fnana.2014.00132

Quantitative relationships in delphinid neocortex

Heidi S. Mortensen 1,2 , Bente Pakkenberg 1 , Maria Dam 2 , Rune Dietz 3 , Christian Sonne 3 ,
Bjarni Mikkelsen 4 and Nina Eriksen 1*
1
Research Laboratory for Stereology and Neuroscience, Bispebjerg and Frederiksberg University Hospitals, Copenhagen, Denmark
2
Research Department, Environment Agency, Torshavn, Faroe Islands
3
Department of Bioscience, Institute for Bioscience - Arctic Research Centre, Roskilde, University of Aarhus, Roskilde, Denmark
4
Museum of Natural History, Torshavn, Faroe Islands

Edited by: Possessing large brains and complex behavioral patterns, cetaceans are believed to be
Patrick R. Hof, Mount Sinai School highly intelligent. Their brains, which are the largest in the Animal Kingdom and have
of Medicine, USA
enormous gyrification compared with terrestrial mammals, have long been of scientific
Reviewed by:
interest. Few studies, however, report total number of brain cells in cetaceans, and even
Mary Ann Raghanti, Kent State
University, USA fewer have used unbiased counting methods. In this study, using stereological methods,
Muhammad A. Spocter, Des Moines we estimated the total number of cells in the neocortex of the long-finned pilot whale
University, USA (Globicephala melas) brain. For the first time, we show that a species of dolphin has
*Correspondence: more neocortical neurons than any mammal studied to date including humans. These
Nina Eriksen, Research Laboratory
cell numbers are compared across various mammals with different brain sizes, and the
for Stereology and Neuroscience,
Bispebjerg Hospital, Bispbjerg function of possessing many neurons is discussed. We found that the long-finned pilot
Bakke 23, 2400 whale neocortex has approximately 37.2 × 109 neurons, which is almost twice as many
Copenhagen, Denmark as humans, and 127 × 109 glial cells. Thus, the absolute number of neurons in the
e-mail: [email protected]
human neocortex is not correlated with the superior cognitive abilities of humans (at least
compared to cetaceans) as has previously been hypothesized. However, as neuron density
in long-finned pilot whales is lower than that in humans, their higher cell number appears
to be due to their larger brain. Accordingly, our findings make an important contribution to
the ongoing debate over quantitative relationships in the mammalian brain.

Keywords: neocortical cell number, glia/neuron ratio, neocortical cell density, animal cognition, stereology

INTRODUCTION Cytoarchitectural organization is very complex (Hof et al.,

Cetaceans are divided into odontocetes (toothed whales) and 2005), and does not resemble that of terrestrial mammals; layer
mysticetes (baleen whales). These two suborders appeared and I is far more cellular, layer II contains atypical neurons, and layer
began to diverge in the early Oligocene, about 30 million III contains very large pyramidal neurons (Glezer, 2002; Hof et al.,
years ago (Gingerich et al., 1983). During secondary adapta- 2005). Cortical layer IV is absent or very poorly developed, and
tion to water, cetaceans underwent major transformations in so inputs, outputs, and interneuronal connections are very differ-
body form and physiology, resulting in large, highly encephalised, ent than those in other mammals (Glezer et al., 1988; Morgane
and extremely gyrified brains compared to those of terrestrial and Glezer, 1990). The cortical arrangement of functional areas
mammals (Oelschläger and Oelschläger, 2002; Marino, 2008). has also changed. The frontal region in the cetacean brain is very
This is especially true for odontocetes such as the sperm whale modest or even absent (Morgane et al., 1980). Compared to pri-
(Physeter macrocephalus) (∼10 kg), which has the largest brain mates, it displays its own unique pattern of differentiation, but
in the Animal Kingdom, but it is not the largest animal alive it is distinctly laminated and comprises several cortical fields as
(Marino, 2004). The cetacean brain differs from the terrestrial in other lobes (Hof et al., 2005). Electrophysiological mapping
mammalian brain in many ways. It has a very high level of gyri- studies have placed both the auditory and visual cortices in the
fication, and like the rest of the whale body, it possesses several parietal regions of dolphins and porpoises (Supin et al., 1978),
traits that indicate adaptation to water. The migration of the whereas they are located in the temporal (auditory) and occipi-
blowhole from the front to the top of the head facilitates breath- tal (visual) regions in terrestrial mammals. The auditory region
ing at the water’s surface. This migration has also changed the is located on the suprasylvian gyrus, which adjoins the visual
shape of the brain and the cranial nerve distribution (Oelschläger areas in the lateral gyrus and dorsal parietal regions. There is
and Oelschläger, 2009). Cetaceans have been observed to be the no intervening cortex between the auditory areas and the visual
most gyrencephalic mammals studied to date, also more than areas or between the visual-auditory areas and the sensorimotor
predicted based on comparison with other mammals. This could areas (Glezer et al., 1988). The auditory systems are smaller in
indicate a morphological alteration of the telencephalon associ- mysticetes than in odontocetes, but the mysticete visual system
ated with the return to the marine environment (Manger et al., contains more axons (Oelschläger and Oelschläger, 2002). The
2012). olfactory system is greatly reduced, and in fact is almost absent.

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Mortensen et al. Quantitative relationships in delphinid neocortex

This is more obvious in odontocetes than mysticetes (Oelschläger (Baird et al., 2002), and they do not usually dive as deep as other
and Oelschläger, 2002). pelagic species such as beaked whales or sperm whales (Heide-
Odontocetes also show specialized hemispheric independency, Jørgensen et al., 2002). Echolocation is used for navigation and
such as independent eye movements and closure in beluga whales fouraging, and sounds of the long-finned pilot whale range from
(Delhinapterus leucas) and unihemispheric sleep in Amazon to 3–18 KHz (Busnel and Dziedzic, 1966). Audiogram shows best
river dolphins (Inia geoffrensis) (Mukhametov, 1987), beluga hearing from 11 to 50 KHz (Pacini et al., 2010). Like other del-
whales (Lyamin et al., 2002), and bottlenose dolphins (Tursiops phinids, the long-finned pilot whale seems to see well in both air
truncatus) (Mukhametov et al., 1988; Ridgway et al., 2006). and water (Herman et al., 1975; Waller, 1992).
The Encephalization Quotient (EQ) is a measure of observed Using the optical fractionator, we estimated the total number
brain size relative to expected brain size (Jerison, 1985). of neocortical cells in the long-finned pilot whale, and found that
Odontocetes brains are highly encephalized and significantly long-finned pilot whales have the highest number of neocorti-
larger than expected for body size. The EQ of living odonto- cal cells of any species studied to date, including humans. These
cetes are generally on par with non-human primates. But some numbers are discussed. We also quantified the auditory and visual
odontocetes species belonging to the Delphinidae family contains cortices (both cell numbers and volume), and compared them to
several species with exceptional high EQs above 4.0, a level of other mammals with different brain and body sizes.
encephalization second only to humans (EQ ∼7) (Marino, 2004).
This had led to speculations that the large brains of cetaceans MATERIALS AND METHODS
could be related to cognitive demands associated with echolo- Brains from 10 long-finned pilot whales from the Faroe Islands
cation (Jerison, 1985; Ridgway and Au, 1999) or a response to were used in this study (Figures 1A,B). The brains were collected
social forces, because cetaceans display complex social patterns as part of a local hunt overseen by local authorities and the North
(Payne and Mcvay, 1971; Rendell and Whitehead, 2001) and Atlantic Marine Mammal Commission (NAMMCO). All animals
behaviors such as self-recognition (Delfour and Marten, 2001; were healthy and classified as juveniles or adults depending on
Reiss and Marino, 2001), cooperation, and tool use (Krutzen sexual maturity (Desportes et al., 1993; Martin and Rothery,
et al., 2005). These capabilities, however, are also possessed by 1993). Brains were kept in 4.5% formalin for 4 months. A single
other animals including great apes (Roth and Dicke, 2005), ele- hemisphere from each animal was chosen at random for analy-
phants (Roth and Dicke, 2005), and even some birds (Emery, sis. Characteristics of individual animals are shown in Table 1.
2006). Yet, other studies show the opposite; that the large cetacean We were interested in three different brain regions; the entire
brain is merely an efficient thermogenetic organ that effec- neocortex, the primary auditory cortex and the primary visual
tively counteracts heat loss to the water (Manger, 2006), and cortex.
that cetacean intelligence is qualitatively not different to other
vertebrates (Manger, 2013; Patzke et al., 2013). Thus, it has CORTICAL MAPPING
recently been argued that mammalian brains should not be scaled The cytoarchitecture of the primary visual and primary auditory
equally, as brains of the same size do not always contain simi- cortices resembled that found by Furutani in the striped dol-
lar cell numbers (Herculano-Houzel, 2011b). Rather, the absolute phin (Stenella coeruleoalba), the Risso dolphin (Grampus griseus),
number of neurons, irrespective of brain or body size, may and the bottlenose dolphin (Furutani, 2008) and harbor por-
be a better predictor of cognitive abilities (Herculano-Houzel, poise (Walloe et al., 2010) (see, Figures 2A–C for cytoarchitecture
2011a). in the three brain regions of interest in the long-finned pilot
Previous studies report estimations of the total number whale brain). Figures 2A–C shows that there is a difference in
of brain cells in both mysticetes [common Minke whale laminar pattern between the entire neocortex (Figure 2A), the
(Balaenoptera acutorostrata) (Eriksen and Pakkenberg, 2007)] primary auditory (Figure 2B), and visual cortices (Figure 2C).
and odontocetes [harbor porpoise (Phocoena phocoena) (Walloe However, it was not possible to identify the subdivisions of
et al., 2010)], but no such estimation has been performed for neocortex using distinct cytoarchitectural features as described
delphinid species, which are expected to have high numbers of by Hof and Van Der Gucht (2007) in the humpback whale
neocortical neurons due to high EQ levels and advanced cognitive (Megaptera novaeangliae) or the presence of well-visible cellu-
abilities. Moreover, neither cetaceans nor any other large-brained lar modules as seen in layer II in the occipital lobe of the
species have been found to possess more neocortical neurons humpback whale. We therefore, relied on macroscopic iden-
than humans. In this study, we studied the brain from the long- tification of primary auditory and visual cortices. Thus, the
finned pilot whale (Globicephala melas Traill, 1809), which has cortical structure of the long-finned pilot whale was assumed
an EQ of 2.39 (Marino, 2004). The long-finned pilot whale is a to follow that of other odontocetes (e.g., bottlenose dolphin,
delphinid around 5–6 m long (Desportes et al., 1993) that lives harbor porpoise), with primary auditory cortex on the supra-
in large matriarchal pods (Ottensmeyer and Whitehead, 2003). sylvian gyrus, between the suprasylvian sulcus and the lateral
It inhabits the deep waters of the North Atlantic ranging from sulcus, and primary visual cortex on the lateral gyrus between
the North Atlantic Ocean from Ungava Bay, Disko in Western the lateral sulcus and entolateral sulcus in the parietal region
Greenland, 68◦ N in Eastern Greenland, Iceland, the Faroe Islands, (Ladygina et al., 1978; Supin et al., 1978; Morgane and Glezer,
and Norway, south to North Carolina, the Azores, Madeira, and 1990). These two cortical areas were marked on the dural surface
Mauritania, including the Western Mediterranean (Rice, 1998). by different colored tissue dye (CANCER Diagnostics) for later
They feed on squid and other prey normally found down to 600 m identification.

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Mortensen et al. Quantitative relationships in delphinid neocortex

FIGURE 1 | The long-finned pilot whale (Globicephala melas) and its brain. (A) Image of a long-finned pilot whale swimming near the Faroese Islands
(copyright Farophoto Inc.). (B) Representative image of a long-finned pilot whale brain.

Table 1 | Characteristics of individual animals used in this study. in approximately 4–10 bars per slab (Figure 3B). The subsam-
pled bars were embedded in historesin (KULZER). Shrinkage was
Individual Hemisphere Sex Length (m) Sexual Total brain
less than 5% and therefore, was negligible (Dorph-Petersen et al.,
maturity weight (g)
2000). From each bar, a 40 µm-thick section was cut and stained
1 Right M 3.2 Juvenile 3062 with a modified Giemsa stain. After sampling had been performed
2 Left F 2.7 Juvenile 3142 on the entire neocortex, the primary auditory and visual cortices
3 Left M 3.4 Juvenile 3272 were processed in the same manner.
4 Left M 3.0 Juvenile 3487 The optical disector, a 3D probe, was placed uniformly ran-
5 Left M 4.6 Juvenile 3705 domly on each the sections. Counting frames were superimposed
6 Left M 4.3 Juvenile 3858 on the magnified image of the tissue, and counting took place
7 Right F 4.9 Adult 3473 inside the thick sections using an unbiased counting frame.
8 Left F 4.8 Adult 3665 Different counting frames were used consecutively for neurons
9 Left M 5.8 Adult 3800 (counting frame area 4500 µm2 ) and glial cells (counting frame
10 Left M 5.6 Adult 4618 area 1200 µm2 ), due to higher numbers of glial cells. All cells were
counted in a microscope array with a BX-50 Olympus microscope
with a 100× oil immersion objective (2000× final magnification)
STEREOLOGICAL CELL COUNTING a motorized x-y stage, an electronic microcator (Heidenhain),
Single hemispheres were cut into 1 cm-thick slabs and embedded and a computer running CAST-GRID software (Visiopharm,
in agar (Figure 3A). The volume of the three brain regions (Vref ) Hørsholm, Denmark). Cell density was equal to the number of
was calculated using a point-counting grid and the following cells counted in all disectors divided by the volume in which they
equation (Gundersen and Jensen, 1987): were counted (Gundersen et al., 1988):
   
Vref = t • a p • P Q
Nv =   
a frame • hdis • P
where Vref is the total volume of each brain region, t is the thick- 
ness of the slab, a(p) where NV is the cell density, Q is the number of cells counted,
 is the predetermined and constant area per
point (mm2 ), and P is the number of points hitting the region a(frame) is the area of the counting frame, hdis is the disec-
of interest (ROI). A point-counting grid was randomly placed tor height. The disector height was 20 µm for both cell types.
on each slap, and all points hitting the ROI were counted. The Total cell number was found by multiplying Vref by Nv . The
average number of slabs was 18.1 (range: 16–20), and the aver- mean number of counted cells per specimen was 198 for neu-
age number of points hitting the entire neocortex was 150 (range: rons (range: 145–230) and 238 for glial cells (range: 165–481) for
130–163). The average number of points hitting the auditory cor- entire neocortex; auditory cortex neurons 176 (range: 141–229),
tex was 168 (range: 142–201), and 138 (range: 129–150) for the and glial cells 185 (range: 133–234); visual cortex neurons 202
visual cortex. (range: 147–265), and glial cells 204 (range: 133–247).
A combination of Cavalieri’s principle and the optical disector
technique (Gundersen and Jensen, 1987; West et al., 1991) was COUNTING CRITERIA
applied to cell counting. The cells counted were neurons and glial Cells were counted in all layers of cortex from the pial surface
cells. From the brain slabs neocortical bars (4 mm) were subsam- to the gray/white transition (Figures 2A–C). Both neurons and
pled from the entire neocortex using systematic uniform random glial cells were estimated in the selected brain regions, but we did
sampling (SURS) (Pakkenberg and Gundersen, 1997), resulting not distinguish between different types of neurons or glial cells.

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Mortensen et al. Quantitative relationships in delphinid neocortex

FIGURE 2 | Anatomy of the long-finned pilot whale brain. Laminar auditory cortex and (C) the primary visual cortex. (D) Different brain
organization in the three regions of interest (not the especially cells. Note the large difference in size of neurons (black arrow) and
difference in cell density) in (A) the entire neocortex, (B) the primary glial cells (white arrow).

Neurons were identified as having a clearly defined nucleus with a SURS variance for the sum of areas is the uncertainty of
pale surrounding cytoplasm and a dark, centrally located nucleo- sampling between sections, because repeated estimates based on
lus. Glial cells were identified by their smaller size and their lack of different sections may vary:
cytoplasm. The nucleolus was easily identified in most of the glial
cells, and the nucleolus was chosen as the counting item for both 3 (Pi • Pi − Noise) − 4 (Pi • Pi+1 ) + (Pi • Pi+2 )
VarSURS  a =
neurons and glial cells (Figure 2D). If a nucleolus could not be 240
identified in neurons, the nucleus was used as the counting item.
where Pi is the number of points counted on one section, Pi+1
If the cell had more than one nucleolus the most centrally located
is the number of points counted on the next section. The total
was used as counting item.
sampling variance, CE(P), is estimated from:
STATISTICAL ANALYSIS   VARNoise + VARSURS  a
The precision of each estimate was determined by the coeffi- CE P = 
cient of error CE (Gundersen and Jensen, 1987). CE is a function P
of the noise effect, also known as the point
 counting variance, Differences between individuals were calculated as the coefficient
and the SURS variance for sums of areas, a (Gundersen et al.,
of variation (CV) using this formula:
1999). Thenoise effect is the uncertainty that comes from point
counting, P: SD
CV =
 mean
b 
Noise = 0.0724 • √ • n• P As a rule CE is considered optimal, when it is approximately
a
one-half or less of the observed inter-individual variance (CVobs ):
√
where n is the number of sections, b/ a is the average profile
CE2
shape (found from eyeballing the nomogram from Gundersen
2
≤ 0.5
and Jensen (1987). CVobs

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Mortensen et al. Quantitative relationships in delphinid neocortex

Table 2 | Estimated cell numbers and density in the long-finned pilot

whale brain.

Neocortex CE (CV) Auditory CE (CV) Visual CE (CV)

cortex cortex

MEAN NEURON NUMBER, 109

F 36.9 0.08 (0.04) 2.2 0.08 (0.05) 2.1 0.08 (0.18)
M 37.4 0.08 (0.17) 2.4 0.08 (0.25) 2.4 0.08 (0.17)
All 37.2 0.08 (0.14) 2.3 0.08 (0.20) 2.3 0.08 (0.18)
MEAN GLIAL CELL NUMBER
F 124 0.07 (0.23) 8.6 0.08 (0.18) 7.6 0.07 (0.13)
M 128 0.08 (0.21) 8.2 0.08 (0.29) 7.6 0.08 (0.09)
All 127 0.07 (0.20) 8.3 0.08 (0.24) 7.6 0.08 (0.10)
MEAN DENSITY OF NEURONS, 106 /cm3
F 31.5 25.8 29.8
M 30.8 28.4 34.1
All 30.9 27.4 32.3
MEAN DENSITY OF GLIAL CELLS, 106 /cm3
F 104 102 109
M 105 94.6 106
All 105 97.3 107
MEAN VOLUME, cm3
F 1178 0.04 (0.09) 84 0.03 (0.07) 70 0.04 (0.05)
M 1222 0.04 (0.14) 85 0.03 (0.16) 71 0.04 (0.05)
All 1209 0.04 (0.12) 85 0.03 (0.13) 71 0.04 (0.05)
GLIAL CELL/NEURON RATIO
All 3.4/1 3.6/1 3.4/1
FIGURE 3 | Sampling procedure. (A) After coloring and embedding in
agar, the brain was coronally cut into 1 cm thick consecutive slabs.
(B) Tissue wedges were sampled from every second slab using SURS.
Each wedge was cut into bars, leaving approximately 40 bars. From these
40 bars, 10 bars were subsampled from each region (neocortex, auditory
cortex, visual cortex), and embedded in historesin.

Then the variance is dominated by the biological variance

between subjects.
Because data were not normally distributed, Mann–Whitney
U-tests were used to test for sex differences in cell number, cell
density, and neocortical volume. We also wished to investigate
the sexual dimorphism within the maturity groups (juvenile or
adult), and for this purpose we used a Kruskal–Wallis equality-
of-populations rank test. Spearman Rank Sum Correlations were
used to assess relationships among body length, neocortical vol-
ume, number of cells, and brain weight. Wilcoxon’s Sign Rank test
was used to test for differences between volumes of auditory and
visual cortices. Statistical analysis was performed using Stata 12.1 FIGURE 4 | Neocortical cell number in individual long-finned pilot
whales. Neuron numbers range from 29.1–46.3 × 109 , and glial cells range
software (StataCorp LP, USA). Statistical significance was set at
from 99.9–183 × 109 . Diamonds indicate neurons, circles indicate glial cells.
P < 0.05. Abbreviations: AF, adult females (pink), AM, adult males (green), JF,
juvenile females (dark red), JM, juvenile males (blue).
RESULTS
Table 2 summarizes the mean cell numbers found in the long-
finned pilot whales, and Figure 4 shows the number of neurons individuals (range: 99.9–183 × 109 ), whereas the neuron num-
and glial cells in the entire neocortex in each individual ani- ber was less variable (range: 29.1–46.3 ×109 ). It was estimated
mal. We found that, on average, long-finned pilot whales possess that long-finned pilot whales have an average of 2.3 × 109 neu-
37.2 × 109 neocortical neurons and 127 × 109 neocortical glial rons and 8.3 × 109 glial cells in the auditory cortex, and 2.3 ×
cells, with a glial cell to neuron ratio of 3.4/1. Figure 4 shows 109 neurons and 7.6 × 109 glial cells in the visual cortex. Next,
that the glial cell number was quite variable among the ten we compared total neocortical cell numbers across different

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Mortensen et al. Quantitative relationships in delphinid neocortex

mammals (Figures 5A,B) and cell numbers in two functional cor- volume and total glial cell number. No significant correlations
tices across marine mammals (Figures 5C,D) based on existing were found between brain weight and cell density or between
stereological literature. We found that the long-finned pilot whale total neocortical volume and cell density. The volume of the audi-
has the highest number of neocortical cells estimated to date. In tory cortex was significantly greater than that of the visual cortex
particular, the long-finned pilot whale has almost twice as many (P = 0.018, Wilcoxon), but there were no significant differences
neocortical neurons as humans (Pakkenberg and Gundersen, in neuron number or glial cell numbers between the auditory and
1997). Long-finned pilot whales also have the highest number of visual cortices.
neocortical cells in both auditory and visual cortices compared Finally, across mammals, we compared actual and expected
with other marine mammals. neocortical neuron numbers as a function of body weight
Furthermore, we found that long-finned pilot whales have a (Figure 6A) and brain weight (Figure 6B) based on existing
mean neocortical neuron density of 30.9 × 106 /cm3 and a mean stereological literature (Pakkenberg and Gundersen, 1997; Jelsing
neocortical glial cell density of 105 × 106 /cm3 . Mean neuron den- et al., 2006; Christensen et al., 2007; Eriksen and Pakkenberg,
sity in the auditory cortex is 27.4 × 106 /cm3 and mean neuron 2007; Walloe et al., 2010). We found that the number of neocor-
density in the visual cortex is 32.3 × 106 /cm3 . Mean glial cell den- tical neurons in long-finned pilot whales exceeds the expected
sity in the auditory cortex is 97.3 × 106 /cm3 and in the visual value with respect to body weight, although not to the same
cortex is 107 × 106 /cm3 . extent as harbor porpoises or humans. The number of neocortical
There was no sexual dimorphism in cell number, neocorti- neurons in long-finned pilot whales is just below the expected
cal volume, or neuron density and no significant correlations value with respect to brain weight.
between brain weight and total cell number or between neocor-
tical volume and total neuron number. Likewise, there was no DISCUSSION
sexual dimorphism within the two maturity groups. There was no The question of dolphin “intelligence” has long been a subject of
correlation (r2 = 0.6, P = 0.08, Spearman) between neocortical intense dispute and speculation, ranging from early affirmations

FIGURE 5 | Comparative analysis of the long-finned pilot whale neocortex. many neurons as humans, as well as (B) the highest number of neocortical
(A) The long-finned pilot whale is estimated to have the highest number of glial cells. Similar findings were observed for the (C) primary auditory cortex
neocortical neurons than any mammal studied to date, with almost twice as and (D) visual cortex. Diamonds indicate neurons, circles indicate glial cels.

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Mortensen et al. Quantitative relationships in delphinid neocortex

FIGURE 6 | Expected neuron numbers. (A) Long-finned pilot whales 95% confidence interval. References: Minke whale (Eriksen and
have a higher than expected number of neocortical neurons relative to Pakkenberg, 2007), harbor porpoise and harp seal (Walloe et al., 2010),
body weight, although not as high as that for humans or harbor domestic pig (Jelsing et al., 2006), rat (Korbo et al., 1990), Rhesus
porpoises, and (B) a slightly lower than expected number of monkey (macaque) (Christensen et al., 2007), human (Pakkenberg and
neocortical neurons relative to brain weight. Gray shade resembles Gundersen, 1997).

(Lilly and Miller, 1961; Lilly, 1965) to recent denial (Manger, (including three species of dolphins) (Delfour and Marten, 2001;
2006, 2013). Across species, neuron number is widely considered Reiss and Marino, 2001; Roth and Dicke, 2005; Plotnik et al.,
to be a major determinant of neural function and, as a con- 2006) as well as in magpies (Prior et al., 2008), and the use of tools
sequence, behavior (Williams and Herrup, 1988). Neurons are has been observed in dolphins (Krutzen et al., 2005), great apes,
assumed to be somehow responsible for the evolution of intel- elephants (Roth and Dicke, 2005), and birds (Roth and Dicke,
ligence, as species with larger brains (and thus, more neurons) 2005). The bird brain, however, is structurally very different from
generally show a greater range and versatility of behavior than the mammalian brain (Reiner et al., 2005), and the number
species with smaller brains (Jerison, 1985; Marino, 2002). of neurons in birds is much less than that found in mammals
In this study, we show that the long-finned pilot whale, (unpublished data). Across animal classes, therefore, the num-
a species of dolphin, has twice as many neocortical neurons ber of neurons is not equal to cognitive capability; rather, these
as humans. As neocortical neurons account for 90–95% of all capabilities appear to be cases of convergent evolution (Emery,
neurons in the cerebrum (Karlsen and Pakkenberg, 2011), this 2006).
number is approximately equivalent to absolute neuron number. The cognitive capabilities of the long-finned pilot whale (and
According to the theory that the absolute number of neurons most other cetaceans for that matter) are unknown, but there
predicts cognitive superiority (Herculano-Houzel, 2011a), the have been extensive studies of captive bottlenose dolphins. Most
long-finned pilot whale should be cognitively superior to all research has revolved around acoustical behavior, with evi-
other species studied, including humans. However, long-finned dence that bottlenose dolphins are capable of understanding
pilot whales show a higher than expected number of neocortical and manipulation of symbols, interpretation of images (Herman,
neurons relative to body weight, but not to the same degree as 2010), social perception (as described above; e.g., mirror recogni-
humans or harbor porpoises. Still, this is an unusually high num- tion), and numerical cognition (Jaakkola et al., 2005). These skills
ber of neurons compared to what have been reported recently have been referred to some level of mental capacity or aspect of
in other large-brained animals using non-stereological methods intelligence (Herman, 2010). However, this has been disputed by
(African elephants: 5.6 × 109 , gorillas: 9 × 109 , orangutans; Manger, especially since dolphins lack stage 6 (the highest stage)
8 × 109 ) (Herculano-Houzel and Kaas, 2011; Herculano-Houzel understanding of object performance (Manger, 2013).
et al., 2014). The neocortical neuron number of the bottlenose dolphin is
unknown, but both long-finned pilot whales and harbor por-
COGNITIVE BEHAVIOR ACROSS SPECIES poises have an unexpected high number of neocortical neurons,
An animal’s cognitive capability is widely used as an indicator of harbor porpoises even to same degree as humans (Figure 6).
mental capacity or intelligence (Roth and Dicke, 2005). Advanced It is not known whether this high neuron number implies
cognitive capabilities are observed in a variety of animals. For cognitive and/or functional impact, but it is tempting to spec-
example, social transmission of complex songs has been observed ulate that these numbers may be related to some advanced
in both birds and mysticetes (Payne and Mcvay, 1971; Mundinger, cognitive behaviors (here assuming that some parallels can be
1980), self-recognition has been observed in several mammals drawn among delphinid species). However, these behaviors are

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Mortensen et al. Quantitative relationships in delphinid neocortex

also seen separately in other mammals with a lower neuron auditory cortex volume was larger than visual cortex volume.
number. We also found that long-finned pilot whales have twice as many
neurons in the auditory and visual cortices as harbor porpoises
GLIA/NEURON RATIO (Walloe et al., 2010). Since the density of neurons are around three
In addition to a high number of neocortical neurons, long-finned times lower than in harbor porpoises (neurons: 98.7 × 106 /cm3 ,
pilot whales also have the highest number of neocortical glial unpublished data), the larger number of neurons in the two cor-
cells in any species studied to date. The glia/neuron ratio of tices in the long-finned pilot whale are most likely due to larger
3.4/1 observed here is lower than that observed in Minke whales volume. Surprisingly, however, we found no difference in neu-
(7.7/1) (Eriksen and Pakkenberg, 2007), whereas it is higher ron number between the primary auditory and visual cortices in
than that in humans (1.4/1) (Pakkenberg and Gundersen, 1997), the long-finned pilot whale, even though delphinids rely heavily
and harbor porpoises (2.4/1) (Walloe et al., 2010; Eriksen and on audition. Similar results were found by Poth and colleagues,
Pakkenberg, 2014). Glia/neuron ratios have not been reported for who estimated neuron number per neocortical unit in sensori-
other cetaceans with the exception of bottlenose dolphins (∼3/1) motor, auditory, and visual cortices in six species of odontocetes
and fin whales (Balaenoptera physalus) (∼5/1) (Oelschläger and (Poth et al., 2005). Thus, auditory specialization might lie else-
Oelschläger, 2002), for which cell counts were not stereologically where in the brain, or odontocetes may rely more heavily on
estimated. Still, these numbers indicate that there is a tendency vision than previously assumed (Teilmann et al., 2007; Herman,
for increased glia/neuron ratios with greater brain mass. Neurons 2010). However, our finding, though interpreted with caution,
are increasingly energetically expensive in larger neocortices, and might suggest that the high number of neocortical neurons in
an increased number of glial cells might proliferate to provide the long-finned pilot whale is probably not a product of cog-
metabolic support to neurons. Thus, the absolute number of glial nitive demands associated with echolocation, as was previously
cells in the brain provides an indication of the metabolic demand suggested (Jerison, 1985; Ridgway and Au, 1999).
of neighboring neurons (Sherwood et al., 2006). Minke whales For comparison, using non-stereological methods, it has
also have very large neuronal perikaryon volume, suggesting that been estimated that macaques (Macaca mulatta) with a brain
large neurons may require a large number of glial cells (Eriksen weight of ∼80 g have 5.4 × 106 neurons in the auditory cortex,
and Pakkenberg, 2007, 2014). As glia/neuron ratio may reflect and baboons (Papio cynocephalus Anubis) with a brain weight
the importance of glia in facilitating neuron growth and hence, of ∼150 g have 7.1 × 106 neurons in the auditory cortex (Wong
neocortical function, it likely plays an important role in cetaceans et al., 2013).
(Eriksen and Pakkenberg, 2007). One other study reports the cell density in the visual cortex
It has been hypothesized recently that an unusually high num- of the bottlenose dolphin (Garey and Leuba, 1986), and the neu-
ber of glial cells together with unihemispheric sleep is an effective ron density (69.7 × 103 /mm3 ) is around twice that of long-finned
way to counteract heat loss in cetaceans (Manger, 2006). This pilot whale in this study. However, both anterior and posterior
would especially apply to deep diving species. As mentioned end of the lateral gyrus was only reported for a single adult
above all cetaceans studied have shown a very high number of glial animal, and thus it is hard to compare to our results.
cells in their neocortex (Eriksen and Pakkenberg, 2007; Walloe In conclusion, we found that the neocortex of the long-
et al., 2010) with the highest glia/neuron ratio is observed in finned pilot whale contains substantially more neurons and glial
Minke whales (Eriksen and Pakkenberg, 2007). However, Minke cells than the neocortex of other large-brained species includ-
whales also have rather large corpus callosi (Ratner et al., 2010), ing humans. As expected, due to their larger brains, odontocetes
much larger than investigated in odontocete species (Tarpley have substantially more brain cells than mysticetes (Marino, 2002,
and Ridgway, 1994; Keogh and Ridgway, 2008), indicating that 2004; Marino et al., 2007). Our results underscore that corre-
the specialized hemispheric independency is probably absent in lations between cognitive performance and absolute neocortical
mysticetes (Ratner et al., 2010). neuron numbers across animal orders or classes are of limited
value, and attempts to quantify the mental capacity of a dolphin
FUNCTIONAL CORTICES for cross-species comparisons are bound to be controversial.
Extensive work has been performed on cytoarchitecture in func-
tional corticies (limbic lobe, visual cortex) in a number of odon- REFERENCES
Baird, R. W., Borsani, J. F., Hanson, M. B., and Tyack, P. L. (2002). Diving and
tocetes by Glezer, Morgan and Garey and co-workers (Morgane night-time behavior of long-finned pilot whales in the Ligurian Sea. Mar. Ecol.
et al., 1982; Garey and Leuba, 1986; Glezer and Morgane, 1990; Prog. Ser. 237, 301–305. doi: 10.3354/meps237301
Glezer et al., 1993). However, only a couple of studies report Busnel, R.-G., and Dziedzic, A. (1966). “Acoustic signals of the pilot whale
actual cell numbers (Poth et al., 2005; Eriksen and Pakkenberg, Globicephala melaena and of the porpoises Delphinus delphis and Phocoena
phocoena,” in Whales, Dolphins and Porpoises, ed K. S. Norris (Berkeley; Los
2007; Walloe et al., 2010). We are well aware that it is a major Angeles, CA: University of California Press), 607–646.
limitation that the functional cortices in this study are not delin- Christensen, J. R., Larsen, K. B., Lisanby, S. H., Scalia, J., Arango, V., Dwork, A.
eated using distinct cytoarchitectual indicators. The cell numbers J., et al. (2007). Neocortical and hippocampal neuron and glial cell numbers
and volume of these cortices must thus be evaluated with cau- in the rhesus monkey. Anat. Rec. (Hoboken) 290, 330–340. doi: 10.1002/ar.
tion. However, the outlining on the cortical surface was a copy of 20504
Delfour, F., and Marten, K. (2001). Mirror image processing in three marine
the atlas of the bottlenose dolphin (Ladygina et al., 1978; Supin mammal species: killer whales (Orcinus orca), false killer whales (Pseudorca cras-
et al., 1978; Morgane and Glezer, 1990), which gives it some val- sidens) and California sea lions (Zalophus californianus). Behav. Processes 53,
idation. Consistent with previous studies, we found that primary 181–190. doi: 10.1016/S0376-6357(01)00134-6

Frontiers in Neuroanatomy www.frontiersin.org November 2014 | Volume 8 | Article 132 | 8

Mortensen et al. Quantitative relationships in delphinid neocortex

Desportes, G., Saboureau, M., and Lacroix, A. (1993). Reproductive maturity and Hof, P. R., and Van Der Gucht, E. (2007). Structure of the cerebral cor-
seasonality of male long-finned pilot whales, off the Faroe Islands. Rep. Int. tex of the humpback whale, Megaptera novaeangliae (Cetacea, Mysticeti,
Whal. Commn. (Special Issue 14), 233–262. Balaenopteridae). Anat. Rec. (Hoboken) 290, 1–31. doi: 10.1002/ar.20407
Dorph-Petersen, K. A., Gundersen, H. J., and Jensen, E. B. (2000). Jaakkola, K., Fellner, W., Erb, L., Rodriguez, M., and Guarino, E. (2005).
Non-uniform systematic sampling in stereology. J. Microsc. 200, 148–157. Understanding of the concept of numerically “less” by bottlenose dolphins
doi: 10.1046/j.1365-2818.2000.00750.x (Tursiops truncatus). J. Comp. Psychol. 119, 296–303. doi: 10.1037/0735-
Emery, N. J. (2006). Cognitive ornithology: the evolution of avian intelligence. 7036.119.3.296
Philos. Trans. R. Soc. Lond. B Biol. Sci. 361, 23–43. doi: 10.1098/rstb.2005.1736 Jelsing, J., Nielsen, R., Olsen, A. K., Grand, N., Hemmingsen, R., and Pakkenberg,
Eriksen, N., and Pakkenberg, B. (2007). Total neocortical cell number in the B. (2006). The postnatal development of neocortical neurons and glial cells in
mysticete brain. Anat. Rec. (Hoboken) 290, 83–95. doi: 10.1002/ar.20404 the Gottingen minipig and the domestic pig brain. J. Exp. Biol. 209, 1454–1462.
Eriksen, N., and Pakkenberg, B. (2014). “Comparative stereology studies of brains doi: 10.1242/jeb.02141
from marine mammals,” in Neurostereology, eds P. R. Mouton and A. Iowa (John Jerison, H. J. (1985). Animal Intelligence as encephaliztion. Phil. Trans. R. Soc. B
Wiley & Sons, Inc.), 99–112. 308, 21. doi: 10.1098/rstb.1985.0007
Furutani, R. (2008). Laminar and cytoarchitectonic features of the cerebral cortex Karlsen, A. S., and Pakkenberg, B. (2011). Total numbers of neurons and glial cells
in the Risso’s dolphin (Grampus griseus), striped dolphin (Stenella coeruleoalba), in cortex and basal ganglia of aged brains with Down syndrome–a stereological
and bottlenose dolphin (Tursiops truncatus). J. Anat. 213, 241–248. doi: study. Cereb. Cortex 21, 2519–2524. doi: 10.1093/cercor/bhr033
10.1111/j.1469-7580.2008.00936.x Keogh, M. J., and Ridgway, S. H. (2008). Neuronal fiber composition of the cor-
Garey, L. J., and Leuba, G. (1986). A quantitative study of neuronal and glial pus callosum within some odontocetes. Anat. Rec. (Hoboken) 291, 781–789. doi:
numerical density in the visual cortex of the bottlenose dolphin: evidence for 10.1002/ar.20701
a specialized subarea and changes with age. J. Comp. Neurol. 247, 491–496. doi: Korbo, L., Pakkenberg, B., Ladefoged, O., Gundersen, H. J., Arlien-Soborg, P., and
10.1002/cne.902470408 Pakkenberg, H. (1990). An efficient method for estimating the total number of
Gingerich, P. D., Wells, N. A., Russell, D. E., and Shah, S. M. (1983). Origin of neurons in rat brain cortex. J. Neurosci. Methods 31, 93–100. doi: 10.1016/0165-
whales in epicontinental remnant seas: new evidence from the early eocene of 0270(90)90153-7
Pakistan. Science 220, 403–406. doi: 10.1126/science.220.4595.403 Krutzen, M., Mann, J., Heithaus, M. R., Connor, R. C., Bejder, L., and
Glezer, I. (2002). “Neural Morphology,” in Marine Mammal Biology (An Sherwin, W. B. (2005). Cultural transmission of tool use in bottlenose dol-
Evolutionary Approach), ed R. Hoezel (Oxford, UK: Blackwell Publishing), phins. Proc. Natl. Acad. Sci. U.S.A. 102, 8939–8943. doi: 10.1073/pnas.05002
98–115. 32102
Glezer, I. I., Hof, P. R., Leranth, C., and Morgane, P. J. (1993). Calcium-binding Ladygina, N. M., Mass, A. M., and Supin, A. Y. (1978). Multiple sensory projections
protein-containing neuronal populations in mammalian visual cortex: a com- in the dolphin cerebral cortex. Zh. Vyssh. Nerv. Deiat. 28, 47–1054.
parative study in whales, insectivores, bats, rodents, and primates. Cereb. Cortex Lilly, J. C. (1965). Vocal mimicry in tursiops: ability to match numbers and
3, 249–272. doi: 10.1093/cercor/3.3.249 durations of human vocal bursts. Science 147, 300–301. doi: 10.1126/sci-
Glezer, I. I., Jacobs, M. S., and Morgane, P. J. (1988). Implications of the “initial ence.147.3655.300
brain” concept for brain evolution in Cetacea. Behav. Brain Sci. 11, 75–89. doi: Lilly, J. C., and Miller, A. M. (1961). Vocal exchanges between dolphins: bottlenose
10.1017/S0140525X0005281X dolphins “talk” to each other with whistles, clicks, and a variety of other noises.
Glezer, I. I., and Morgane, P. J. (1990). Ultrastructure of synapses and golgi analysis Science 134, 1873–1876. doi: 10.1126/science.134.3493.1873
of neurons in neocortex of the lateral gyrus (visual cortex) of the dolphin and Lyamin, O. I., Mukhametov, L. M., Siegel, J. M., Nazarenko, E. A., Polyakova, I.
pilot whale. Brain Res. Bull. 24, 401–427. doi: 10.1016/0361-9230(90)90096-I G., and Shpak, O. V. (2002). Unihemispheric slow wave sleep and the state of
Gundersen, H. J., Bendtsen, T. F., Korbo, L., Marcussen, N., Moller, A., Nielsen, K., the eyes in a white whale. Behav. Brain Res. 129, 125–129. doi: 10.1016/S0166-
et al. (1988). Some new, simple and efficient stereological methods and their use 4328(01)00346-1
in pathological research and diagnosis. APMIS 96, 379–394. doi: 10.1111/j.1699- Manger, P. R. (2006). An examination of cetacean brain structure with a novel
0463.1988.tb05320.x hypothesis correlating thermogenesis to the evolution of a big brain. Biol. Rev.
Gundersen, H. J., and Jensen, E. B. (1987). The efficiency of systematic sampling Camb. Philos. Soc. 81, 293–338. doi: 10.1017/S1464793106007019
in stereology and its prediction. J. Microsc. 147, 229–263. doi: 10.1111/j.1365- Manger, P. R. (2013). Questioning the interpretations of behavioral obser-
2818.1987.tb02837.x vations of cetaceans: is there really support for a special intellectual
Gundersen, H. J., Jensen, E. B., Kieu, K., and Nielsen, J. (1999). The efficiency of status for this mammalian order? Neuroscience 250, 664–696. doi:
systematic sampling in stereology–reconsidered. J. Microsc. 193, 199–211. doi: 10.1016/j.neuroscience.2013.07.041
10.1046/j.1365-2818.1999.00457.x Manger, P. R., Prowse, M., Haagensen, M., and Hemingway, J. (2012). Quantitative
Heide-Jørgensen, M. P., Bloch, D., Stefansson, E., Mikkelsen, B., Ofstad, L. H., and analysis of neocortical gyrencephaly in African elephants (Loxodonta africana)
Dietz, R. (2002). Diving behaviour of long-finned pilot whales Globicephala and six species of cetaceans: comparison with other mammals. J. Comp. Neurol.
melas around the Faroe Islands. Wildl. Biol. 8, 307–313. 520, 2430–2439. doi: 10.1002/cne.23046
Herculano-Houzel, S. (2011a). Brains matter, bodies maybe not: the case for exam- Marino, L. (2002). Convergence of complex cognitive abilities in cetaceans
ining neuron numbers irrespective of body size. Ann. N.Y. Acad. Sci. 1225, and primates. Brain Behav. Evol. 59, 21–32. doi: 10.1159/0000
191–199. doi: 10.1111/j.1749-6632.2011.05976.x 63731
Herculano-Houzel, S. (2011b). Not all brains are made the same: new views on Marino, L. (2004). Dolphin cognition. Curr. Biol. 14, R910–R911. doi:
brain scaling in evolution. Brain Behav. Evol. 78, 22–36. doi: 10.1159/000327318 10.1016/j.cub.2004.10.010
Herculano-Houzel, S., Avelino-De-Souza, K., Neves, K., Porfirio, J., Messeder, D., Marino, L. (2008). “Brain evolution,” in Encyclopedia of Marine Mammals, eds W.
Mattos Feijo, L., et al. (2014). The elephant brain in numbers. Front. Neuroanat. F. Perrin, B. Würsig, and J. G. M. Thewissen (San Diego, CA: Academic Press),
8:46. doi: 10.3389/fnana.2014.00046 149–152.
Herculano-Houzel, S., and Kaas, J. H. (2011). Gorilla and orangutan brains con- Marino, L., Connor, R. C., Fordyce, R. E., Herman, L. M., Hof, P. R., Lefebvre, L.,
form to the primate cellular scaling rules: implications for human evolution. et al. (2007). Cetaceans have complex brains for complex cognition. PLoS Biol.
Brain Behav. Evol. 77, 33–44. doi: 10.1159/000322729 5:e139. doi: 10.1371/journal.pbio.0050139
Herman, L. M. (2010). What laboratory research has told us about dolphin Martin, A. R., and Rothery, P. (1993). Reproductive parameters of female
cognition. Int. J.Comp. Psychol. 23, 310–330. longfinned pilot whales (Globicephala melas) around the Faroe Islands. Rep. Int.
Herman, L. M., Peacock, M. F., Yunker, M. P., and Madsen, C. J. (1975). Bottlenosed Whal. Commn. (Special Issue 14), 263–304.
dolphin: duoble-split pupil yields equivalent aerial and underwater diurnal Morgane, P. J., and Glezer, I. I. (1990). “Sensory neocortex in dolphin brain,” in
acuity. Science 1989, 650–659. doi: 10.1126/science.1162351 Sensory Abilities of Cetaceans. Laboratory and Field Evidence, eds J. A. Thomas
Hof, P. R., Chanis, R., and Marino, L. (2005). Cortical complexity in cetacean and R. A. Kastelein (New York, NY: Plenum Press), 101–137.
brains. Anat. Rec. A Discov. Mol. Cell. Evol. Biol. 287, 1142–1152. doi: Morgane, P. J., Jacobs, M. S., and Mcfarland, W. L. (1980). The anatomy of the
10.1002/ar.a.20258 brain of the bottlenose dolphin (Tursiops truncates): surface configurations

Frontiers in Neuroanatomy www.frontiersin.org November 2014 | Volume 8 | Article 132 | 9

Mortensen et al. Quantitative relationships in delphinid neocortex

of the telencephalon of the bottlenose dolphin with comparative anatomical Rice, D. W. (1998). Marine Mammals of the World. Systematics and Distribution.
observation sin four other cetacean species. Brain Res. Bull. 5, 1–107. doi: Lawrence, KS: Society for Marine Mammalogy; Allen Press.
10.1016/0361-9230(80)90272-5 Ridgway, S. H., and Au, W. W. L. (1999). “Hearing and echolocation: dolphin,” in
Morgane, P. J., Mcfarland, W. L., and Jacobs, M. S. (1982). The limbic lobe of the Elsevier’s Encyclopedia of Neuroscience, eds G. Adelmann and B. H. Smith (New
dolphin brain: a quantitative cytoarchitectonic study. J. Hirnforsch. 23, 465–552. York, NY: Elsevier Science BV), 858–862.
Mukhametov, L. M. (1987). Unihemispheric slow-wave sleep in the Amazonian Ridgway, S., Houser, D., Finneran, J., Carder, D., Keogh, M., Van Bonn, W., et al.
dolphin, Inia geoffrensis. Neurosci. Lett. 79, 128–132. doi: 10.1016/0304- (2006). Functional imaging of dolphin brain metabolism and blood flow. J. Exp.
3940(87)90684-7 Biol. 209, 2902–2910. doi: 10.1242/jeb.02348
Mukhametov, L. M., Oleksenko, A. I., and Poliakova, I. G. (1988). [Quantitative Roth, G., and Dicke, U. (2005). Evolution of the brain and intelligence. Trends Cogn.
characteristics of the electrocorticographic sleep stages in bottle-nosed dol- Sci. 9, 250–257. doi: 10.1016/j.tics.2005.03.005
phins]. Neirofiziologiia 20, 532–538. Sherwood, C. C., Stimpson, C. D., Raghanti, M. A., Wildman, D. E., Uddin, M.,
Mundinger, P. C. (1980). Animal cultures and a general theory of cultural evolution. Grossman, L. I., et al. (2006). Evolution of increased glia-neuron ratios in
Ethol. Sociobiol. 1, 183–223. doi: 10.1016/0162-3095(80)90008-4 the human frontal cortex. Proc. Natl. Acad. Sci. U.S.A. 103, 13606–13611. doi:
Oelschläger, H. A., and Oelschläger, J. S. (2009). “Brain,” in Encyclopedia of Marine 10.1073/pnas.0605843103
Mammals, eds W. F. Perrin, B. Würsig, and J. G. M. Thewissen (San Diego, CA: Supin, A. Y., Mukhametov, L. M., Ladygina, T. F., Popov, V. V., Mass, A. M., and
Academic Press), 134–149. Polyakova, I. G. (1978). Electrophysiological Studies of the Dolphin’s Brain (in
Oelschläger, H. H. A., and Oelschläger, J. S. (2002). “Brain,” in Encyclopedia of Russia). (Moscow: Izdatel’stvo Nauka), 7–85.
Marine Mammals, eds W. F. Perrin, B. Würsig, and J. G. M. Thewissen (San Tarpley, R. J., and Ridgway, S. H. (1994). Corpus callosum size in delphinid
Diego, CA: Academic Press), 133–158. cetaceans. Brain Behav. Evol. 44, 156–165. doi: 10.1159/000113587
Ottensmeyer, C. A., and Whitehead, H. (2003). Behavioural evidence for social Teilmann, J., Larsen, F., and Desportes, G. (2007). Time allocation and diving
units in long-finned pilot whales. Can. J. Zool. 81, 1327–1338. doi: 10.1139/ behaviour of harbour porpoises (Phocoena phocoena) in Danish and adjacent
z03-127 waters. J. Cetacean Res. Man. 9, 201–210.
Pacini, A. F., Nachtigall, P. E., Kloepper, L. N., Linnenschmidt, M., Sogorb, A., and Waller, G. N. H. (1992). Ciliary mucscles in the eye of the long-finned pilot whale
Matias, S. (2010). Audiogram of a formerly stranded long-finned pilot whale Globicephala melaena (Traill, 1809). Aquat. Mamm. 18, 36–39.
(Globicephala melas) measured using auditory evoked potentials. J. Exp. Biol. Walloe, S., Eriksen, N., Dabelsteen, T., and Pakkenberg, B. (2010). A neurologi-
213, 3138–3143. doi: 10.1242/jeb.044636 cal comparative study of the harp seal (Pagophilus groenlandicus) and harbor
Pakkenberg, B., and Gundersen, H. J. (1997). Neocortical neuron number in porpoise (Phocoena phocoena) brain. Anat. Rec. (Hoboken) 293, 2129–2135. doi:
humans: effect of sex and age. J. Comp. Neurol. 384, 312–320. 10.1002/ar.21295
Patzke, N., Spocter, M. A., Karlsson, K. A., Bertelsen, M. F., Haagensen, M., West, M. J., Slomianka, L., and Gundersen, H. J. (1991). Unbiased stereolog-
Chawana, R., et al. (2013). In contrast to many other mammals, cetaceans have ical estimation of the total number of neurons in the subdivisions of the
relatively small hippocampi that appear to lack adult neurogenesis. Brain Struct. rat hippocampus using the optical fractionator. Anat. Rec. 231, 482–497. doi:
Funct. doi: 10.1007/s00429-013-0660-1. [Epub ahead of print]. 10.1002/ar.1092310411
Payne, R. S., and Mcvay (1971). Songs of humpback whales. Science 173, 585–597. Williams, R. W., and Herrup, K. (1988). The control of neuron number. Annu. Rev.
doi: 10.1126/science.173.3997.585 Neurosci. 11, 423–453. doi: 10.1146/annurev.ne.11.030188.002231
Plotnik, J. M., De Waal, F. B., and Reiss, D. (2006). Self-recognition in Wong, P., Peebles, J. K., Asplund, C. L., Collins, C. E., Herculano-Houzel, S., and
an Asian elephant. Proc. Natl. Acad. Sci. U.S.A. 103, 17053–17057. doi: Kaas, J. H. (2013). Faster scaling of auditory neurons in cortical areas relative
10.1073/pnas.0608062103 to subcortical structures in primate brains. Brain Behav. Evol. 81, 209–218. doi:
Poth, C., Fung, C., Gunturkun, O., Ridgway, S. H., and Oelschlager, H. H. 10.1159/000350709
(2005). Neuron numbers in sensory cortices of five delphinids compared
to a physeterid, the pygmy sperm whale. Brain Res. Bull. 66, 357–360. doi: Conflict of Interest Statement: The authors declare that the research was con-
10.1016/j.brainresbull.2005.02.001 ducted in the absence of any commercial or financial relationships that could be
Prior, H., Schwarz, A., and Gunturkun, O. (2008). Mirror-induced behavior in construed as a potential conflict of interest.
the magpie (Pica pica): evidence of self-recognition. PLoS Biol. 6:e202. doi:
10.1371/journal.pbio.0060202 Received: 11 September 2014; accepted: 29 October 2014; published online: 26
Ratner, C., Riise, J., Eriksen, N., and Pakkenberg, B. (2010). A postmortem November 2014.
study of the corpus callosum in the common minke whale (Balaenoptera Citation: Mortensen HS, Pakkenberg B, Dam M, Dietz R, Sonne C, Mikkelsen B and
acutorostrata). Mar. Mamm. Sci. 27, 688–700. doi: 10.1111/j.1748-7692.2010. Eriksen N (2014) Quantitative relationships in delphinid neocortex. Front. Neuroanat.
00432.x 8:132. doi: 10.3389/fnana.2014.00132
Reiner, A., Yamamoto, K., and Karten, H. J. (2005). Organization and evolution of This article was submitted to the journal Frontiers in Neuroanatomy.
the avian forebrain. Anat. Rec. A Discov. Mol. Cell. Evol. Biol. 287, 1080–1102. Copyright © 2014 Mortensen, Pakkenberg, Dam, Dietz, Sonne, Mikkelsen and
doi: 10.1002/ar.a.20253 Eriksen. This is an open-access article distributed under the terms of the Creative
Reiss, D., and Marino, L. (2001). Mirror self-recognition in the bottlenose dolphin: Commons Attribution License (CC BY). The use, distribution or reproduction in other
a case of cognitive convergence. Proc. Natl. Acad. Sci. U.S.A. 98, 5937–5942. doi: forums is permitted, provided the original author(s) or licensor are credited and that
10.1073/pnas.101086398 the original publication in this journal is cited, in accordance with accepted academic
Rendell, L., and Whitehead, H. (2001). Culture in whales and dolphins. Behav. practice. No use, distribution or reproduction is permitted which does not comply with
Brain Sci. 24, 309–324. discussion: 324–382. doi: 10.1017/S0140525X0100396X these terms.

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