REPRODUCTIVE

BIOLOGY
OF CARPS

I.J. Singh, Professor

Department of Fishery Biology
College of Fisheries, G.B.P.U.A.&T., Pantnagar
 Onset of 1st maturity and gonadal
recrudescence

Onset of 1st maturity:

 Stage of 1st gonadal development in the life history
of any fish.

Gonadal recrudescence:
 Successive gonadal development after 1st maturity
in subsequent breeding seasons
 Important factors for onset on 1st maturity
2 + years major carps

1- Age 1 + year minor carps

6 months common carp

Natural food level
2-Nutritional status
Artificial feeding

Stimulants

3- Environmental Condition Water quality

Pollution
 Pattern of ovarian development in carps
1- Synchronous type (Synchronise total):
 All oocytes at the same stage of development

2- Group Synchronous type (Synchronise par groups):

 Atleast two populations of oocytes at different
developmental stages.
 Generally spawn once a year and heave relatively a
short breeding season.
 Carps except common carp

3- Asynchronous type (metachrone):

 Oocytes at all stages of development
 Common carp
Pattern of spermatogenesis in teleosts
 Two patterns or types according to the distribution of
spermatogonia.
1- Tubular type or restricted spermatogonial testis:
 Spermatogonia are totally restricted to the distal terminus of the
tubule immediately beneath the tunica albuginea.
 Example - Atheriniformes
2- Lobular type or unrestricted spermatogonial testis:
 Common type
 Any portion of testis shows close relationship between germ cells
at various stages of development and sertoli cells.
 Example - Most of teleost groups including Cypriniformes (carps).
 Neruohormonal regulation of onset of 1st maturity
 Activation of Brain – Pituitary – Gonad (B-P-G) or
Hypothalamo - hypophyseal gonad axis
 Most probably point of initiation in B-P-G axis is gonad.
 Pituitary gland from steroid injected immature fish released
gonadotropin when incubated with hypothalamus or
GnRH/LHRH
 Pituitary gland from immature fish without injection of
steroid did not evoke such response.
 First steroid produced by gonad activates both hypothalamus
and pituitary.
 Subsequently GnRH from hypothalamus stimulates pituitary
for GtH release which in turn regulates steroidogenesis in fish.
 Ultimately processes related with gonadal development move
forward under the concerted regulation by B-P-G axis.
 Brain Hormones
1- Growth Hormone- Releasing hormone (GHRH)

2- Gonadotropin Releasing Hormone (GnRH)

3- Corticotropin – Releasing Factor (CRF)

4- Thyrotropin Releasing Factor (TRF)

5- Gonadotropin Releasing Inhibitory Factor (GRIF)

6- Thyrotropin Release Inhibitory Factor (TRIF)

7- Somatotropin Release Inhibitory Factor (SRIF)

8- Neurohypophyseal Hormones
 Gonadotropin (GtH)
 A dual GtH system has been proposed for several teleosts.
1- One preparation is designated as ConAI or GtH I or
vitellogenic GtH or carbohydrate poor (CP) GtH.
2- Second preparation is designated as ConAII or GtH II or
maturational GtH / steroidogenic GtH or carbohydrate rich
(CR) GtH, adsorbed to Con-A.
 Normally two types of gonadotrops are attributed for above
two GtHs.
 Two types of gonadotrops differing in their location within
PPD and synthetic activity during the reproductive cycle are
identified.
 The synthetic activity of these cell- types vary during the
reproductive development.
 Immunocytochemical localisation of GtH I and
GtH II producing two distinctly different
gonadotrop cell- types in PPD of salmonids.
 Immunoreactive GtH II localised in cells located
mainly in the central regions of the glandular cords
of the PPD.
 GtH I was found in cells located in the periphery of
the glandular cords of the PPD.
 The synthetic activity of these cell- types vary
during the reproductive development.
Two Cell Model for Steroidogenesis
Gonadotropin and Steroidogenesis
 Processes associated with the
gonadal development

1. Testis- Spermatogenesis, spermiogenesis and spermiation in

males

2- Ovary- Oogenesis including vitellogenesis, final oocyte

maturation and ovulation in females.
 Spermatogenesis
 In teleosts, resting single cells in testis lobule or tubule are
referred as primary spermatogonia.
 Primary spermatogonia in the tubule/lobule walls proliferate
to form the clusters of secondary spermatogonia, each cluster
gets enclosed in a cyst.
 The secondary spermatogonia in each cyst divide
synchronously mitotically (212 or 216) to form primary
spermatocyte.
 First meiotic division of primary spermatocyte produces
secondary spermatocyte and 2nd meiotic division produces
haploid spermatids.
 Spermiogenesis
 The metamorphosis of spermatids to spermatozoa is
referred as spermiogenesis.
 Spermiogenesis takes place in the lumen of testis tubule.
 It involves transformation of immotile spermatids into
motile spermatozoa.

Spermiation
 Milt formation involving hydration, change in electrolytes
and pH etc.
 Decrease in density and osmolarity by dilution required
for facilitating release of milt to exterior.
 Hormonal control of steroidogenesis,
Spermatogenesis, and Spermiation
 GtH II controls steroidogenesis in testis.
 It increases levels of of 11-Ketotestosterone and
testosterone in male fish.
 GtH I has no steroidogenic effect.
 11-ketotestosterone is found only in fishes and is
responsible for spermatogonial proliferation
(spermatogenesis).
 Maturational steroid (MS, 17α, 20β-DP ), regulates the
processes of spermiation and milt hydration.
 Oogenesis
 Ovigerous lamellae are the seat for the development of oocyte,
the germ cells or oogonia.
 Two phases of initial growth are recognized-

1- Increase in number by mitotic division

2- Increase in size
 Increase in number takes place by large number of mitotic
division.
 When an oogonium enters the prophase of the 1st meiotic
division it is called primary oocyte.
 After this stage enters 2nd growth phase and vitellogenesis
takes place.
 VITELLOGENESIS
 Primary oocyte passes through two discrete growth phases.
1- Endogenous vitellogenesis:
 Also known as non-vitellogenic which includes deposition of yolk vesicle in
the peripheral region
2- Exogenous vitellogeneis:
 Vitelogenin formation (vitellogenesis) takes place in the liver under the
stimulation of 17β-estradiol produced by ovary.
 17β-estradiol in ovary is produced under the control of steroidogenic GtH
(GtH II).
 Vitellogenin is transported to ovary through blood where it gets
incorporated into ovary through micropinocytosis under the control of
vitellogenic gonadotropin (GtH I)
 Uptaken vitellogenin is deposited in the centre of oocyte as yolk granule.
 Final Oocyte Maturation
 It involves two important developments.

1- Germinal Vesicle Migration (GVM)

2- Germinal Vesicle Break-down (GVBD)

 When vitellogenesis followed by yolk deposition is almost
complete the nucleus migrates (GVM) to the animal pole.
 Nuclear break-down (GVBD) takes place there and 1st meiotic
division is complete forming a secondary oocyte and a polar
body.
 2nd meiotic division which usually takes place after fertilization
in teleosts, converts the secondary oocyte into the ovum and
another polar body.
 Maturation Promoting Factor
(MPF)
 The presence of MS receptors at the oocyte surface is
indicative of the existence of a cytoplasmic factor responsible
for mediation of oocyte maturation.

 This factor is designated as MPF and is possibly produced in

fish oocytes under the control of 17α, 20β-DP.

 MPF activity has been demonstrated in goldfish oocytes

matured by in vivo treatment of HCG.

 Similarly, MPF activity was also detected in oocytes matured

in vitro by 17α, 20β-DP.
 Immature goldfish oocytes matured faster when injected with
MPF extracted from goldfish compared to oocyte induced to
mature in vitro by 17α, 20β-DP.

 In MPF injected oocytes, though GVM did not take place but
the GVBD occurred at the centre.

 MPF activity has been reported to increase before GVBD in

goldfish during in vitro induction of oocyte maturation by
17α, 20β-DP, peaking at 1st meiotic metaphase, decreasing at
1st polar body elimination and increased again and remained
high till insemination.
 MPF is considered a protein consisting of two components,
one catalytic subunit, a homologue of the serine / threonine
protein kinase (cdc2 kinase) and another a regulatory subunit,
cyclin.

 The mature oocytes induced by 17α, 20β-DP were observed to

have 35- KDa inactive and 34-KDa active cdc2 kinase and
cyclinβ.

 The appearance of the 34- KDa active cdc2 kinase coincided

with that of cyclin β just before GVBD.
 It is suggested that 17α, 20β-DP induces synthesis of
cyclin β by oocytes which activates 35- KDa cdc2 kinase to
produce 34- KDa active cdc2 kinase and thus induces
GVBD.
 The available information also indicates that MPF is
similar among vertebrates and invertebrates and it is not
specific.
 It may also be a more general factor responsible for
initiation of breakdown of nuclear membrane and
subsequent cell division.
 OVULATION
 Actual expulsion of oocyte from the follicle.
Follicular separation
 Following final maturation detachment of microvillar
connections between oocyte and follicular layer.
Follicular rupture
 After separation formation of a distinct hole in the follicular
layer through which the oocyte leaves.
 A very specific area of the follicle is weakened and involved in
hole formation at ovulation.
Oocyte expulsion
 Expulsion of oocyte occurs by active contraction of the follicle
with involvement of smooth muscles, pushing the oocyte out.
Mechanism-
Enzymic control- Protease plasmin
Hormonal control-

Plasminogen activator
Plasmin
Plasminogen system
Stimulated by GtH

 Prostaglandins (PG) are involved in ovulation by stimulating

follicular contraction.
 Indomethacin treatment.
 Increase in PG at ovulation.
Migrated Germinal Vesicle and Follicular Separation
 Pituitary-ovarian relay for the stimulation of
final maturation (MS, maturational steroid)
Neurohormonal regulation of final
oocyte maturation and ovulation
Hypothalamus

GnRH GRIF

GnRH
Pituitary
GnRH II
GtH II

Ovary

MS (17α, 20β-DP)
Final maturation and ovulation
Annual changes in Photoperiod, Temperature and Rainfall
(Singh, I.J. and T.P. Singh,Gonado-somatic
1984) index (GSI) in female of Cirrhinus mrigala
(Singh, I.J. and T.P. Singh, 1984)

24

22
20

18
16

14
12
GSI

10
8

6
4

2
0

March April May June July August Sep. Oct. Nov. Dec. Jan. Feb.

Months
 Gonado-somatic index (GSI) in male Cirrhinus mrigala
Gonado-somatic
(Singh, I.J. and T.P. Singh, 1984) index (GSI) in male Cirrhinus mrigala
(Singh, I.J. and T.P. Singh, 1984)

1.8

1.6

1.4

1.2
GSI

0.8

March April May June July August Sep. Oct. Nov. Dec. Jan. Feb.
0.6

0.4

0.2

0
March April May June July August Sep. Oct. Nov. Dec. Jan. Feb.
Months
 Gonado-somatic index (GSI) in female Labeo rohita
(Kumar,Gonado-somatic
A., I. J. Singh and R.N. Ram, 2001) (GSI) in female Labeo rohita
index
(Kumar, A., I. J. Singh and R.N. Ram, 2001)

14

12

10

8
GSI

Ocober Dec. Jan. Feb. March April May June August

0
Ocober Dec. Jan. Feb. March April May June August

Months
 Gonado-somatic index (GSI) in male Labeo rohita
Gonado-somatic(Sinindex
gh, A.K., A.K(GSI)
umar, I.J.Sinin
gh andmale
R.N. Ram,Labeo
2004) rohita
(Singh, A.K., A.Kumar, I.J.Singh and R.N. Ram,2004)
2
2 1.8
1.8 1.6
1.6 1.4
1.4 1.2
1.2 1
1 0.8
GSI

0.8 0.6

0.6 0.4

0.4 0.2

0.2 0
October Nov. Jan. Feb. March April May June August
0
October Nov. Jan. Feb. March April May June August

Months
 Gonado-somatic index (GSI) in male Labeo gonius

2.5

1.5
GSI

0.5

0
Nov. Dec Jan. Feb. March April May June

Months
 Gonado-somatic index (GSI) in female Labeo gonius

4
GSI

0
Nov. Dec. Jan. Feb. March April May June
Months
 Principles of Induced breeding

1. Gonadal development in major carps is completed

in captivity but spontaneous spawning does not
occur due to lack of stimulatory factors and
presence of some inhibitory factors such as excess of
excretory metabolites.

2. Sudden upsurge of GtH is must for spawning in

carps.

3. This is induced to happen with the help of inducing

agents i.e. pituitary extract (PE) or ovaprim and its
alikes.
Levels of external intervention in the hypothalamic-pituitary-ovarian
axis for inducing maturation and ovulation

Hypothalamus Antiestrogens
Dopamine Antagonists
Gonadotropin
Releasing
Hormone (GnRH)

GnRH and
Pituitary Gland GnRH Analogues

Gonadotropin (GtH)
Pituitary Extracts
Ovary GtH Preparations

Oocyte Progestins
Final Maturation Conticosteroids

Ovulation Prostaglandins
Catecholamines
ova
 Principles of Induced Breeding

HYPOPHYSATION
 Use of pituitary gland for induced breeding.
 First priming low dose of PE to females triggers GVM and / or
initiation of GVBD.
 GtH present in PG targets ovary / oocyte directly.
 Second resolving high dose of PE induces GVBD and
ovulation in female fish.
 Low dose of PE to male with resolving dose to female induces
spermiation.
OVAPRIM
 Constituents- SGnRHa (20 μg) and domperidone (10 mg) in
each ml.
 SGnRHa directly acts upon pituitary to induce synthesis
and release of GtH II.
 Domperidone (Dopamine-antagonist) counters negative
impact of dopamine thus facilitating release of more GnRH
and increasing sensitivity of pituitary to GnRH and
SGnRHa.
 As a result release of GtH II from pitutary is increased and
level of MS is elevated causing GVM, GVBD and ovulation
in female and spermiation in male leading to spawning.