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Laboratorio de Toxicologa Acutica, Departamento de Farmacia, Escuela Nacional de Ciencias Biolgicas, IPN. Carpio y Plan de Ayala s/n,
Col. Plutarco Elas Calles Casco de Santo Toms. D.F. CP 11340, Mxico
b
Laboratorio de Histologa, Departamento de Morfologa, Escuela Nacional de Ciencias Biolgicas, IPN. Carpio y Plan de Ayala s/n,
Col. Plutarco Elas Calles Casco de Santo Toms. D.F. CP 11340, Mxico
c
Laboratorio de Biologa Molecular, CIAD Mazatln, Av. Sbalo Cerritos s/n, Mazatln Sinaloa. CP 82010, Mxico
d
Laboratorio de Inmunoqumica I, Departamento de Inmunologa, Escuela Nacional de Ciencias Biolgicas, IPN. Carpio y Plan de Ayala s/n,
Col. Plutarco Elas Calles Casco de Santo Toms. D.F. CP 11340, Mxico
Received 6 April 2006; received in revised form 23 October 2006; accepted 24 October 2006
Available online 16 November 2006
Abstract
Goodeid fish have matrotrophic viviparity, and unlike lecitotrophic fish, yolk loss forces the female to provide the nutritional requirements for
embryonic development. Vitellogenin (VTG) is the yolk precursor protein synthesized in the maternal liver, but there is only circumstantial
evidence regarding VTG supply during the ontogenesis of bony fish with matrotrophic viviparity. Therefore, the goal of the present study was to
identify and quantify VTG during gestation of the black fin goodeid Girardinichthys viviparus and the butterfly split-fin goodeid Ameca
splendens. Females at different gonadic developmental stages were selected in order to evaluate VTG mRNA expression in the maternal liver
using RT-PCR; VTG quantification in maternal muscle and liver, as well as in the embryos, was done using ELISA, and immunohistochemical
detection of VTG was done in the black fin goodeid. The results suggest that VTG supplies nutrients during embryonic development of both
species, which have different life histories. It is possible that the transition from lecitotrophy to matrotrophic viviparity in bony fish with
intraluminal gestation involved adaptive transition strategies that included changes in the relationship between oocytes and follicular cells, as well
as a gradual loss of VTG synthesis during embryonic development.
2006 Elsevier Inc. All rights reserved.
Keywords: Ameca splendens; Embryonic development; Girardinichthys viviparus; Goodeid fish; Matrotrophic viviparity; Vitellogenin
1. Introduction
This paper is part of the 3rd special issue of CBP dedicated to The Face of
Latin American Comparative Biochemistry and Physiology organized by
Marcelo Hermes-Lima (Brazil) and co-edited by Carlos Navas (Brazil), Rene
Beleboni (Brazil), Rodrigo Stabeli (Brazil), Tania Zenteno-Savn (Mexico) and
the editors of CBP. This issue is dedicated to the memory of two exceptional
men, Peter L. Lutz, one of the pioneers of comparative and integrative
physiology, and Cicero Lima, journalist, science lover and Hermes-Lima's dad.
Corresponding author. Tel.: +52 55 5729 6300x62343; fax: +52 55 53 96 35
03.
E-mail address: avegadv@yahoo.com.mx (A. Vega-Lpez).
1095-6433/$ - see front matter 2006 Elsevier Inc. All rights reserved.
doi:10.1016/j.cbpa.2006.10.039
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Fig. 1. Condition factor of females collected at both locations. a) Girardinichthys viviparus from Texcoco Lake (19 25 N; 98 55 W), Estado de
Mexico. b) Ameca splendens from Veneros de Teuchitln (spring from Ameca
river, 20 41 N; 103 43 W), Jalisco, Mexico. W = total mass (g); L = standard
length (mm).
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conventional methods (Ortiz-Ordez et al., in press). Serial 8micron-thick sections were obtained. The slides were then
stained with hematoxylineosin and mounted in synthetic resin
(Entellan) for light microscopy analysis.
3. Results
3.1. Condition factor and reproductive aspects
Gestating females were collected over a year period. A
higher condition factor was observed in the female black fin
goodeid (Fig. 1 a) compared to the female butterfly split-fin
goodeid (Fig. 1 b). Under controlled laboratory conditions,
reproductive information was recorded. In the butterfly split-fin
goodeid, the age of first maturation was 6 months, the gestation
period was 2 months, and the fish gave birth to an average of 21
live offspring with a 40% viability. In the black fin goodeid, the
Table 1
Vitellogenin levels (%) during gestation of Girardinichthys viviparus (Stage
2 = vitellogenesis. Stage 3 = beginning of intraluminal gestation. Stage
4 = embryonic growth. Stage 5 = prenatal embryos)
Target
organ
Stage 2
Stage 3
Stage 4
Stage 5
Parent
muscle
Embryo
Gonad
Liver
0.16 0.2
1.82 1.1
2.03 0.9
1.34 0.7
Table 2
Vitellogenin levels (%) during gestation of Ameca splendens (Stage
2 = vitellogenesis. Stage 3 = beginning of intraluminal gestation. Stage
4 = embryonic growth. Stage 5 = prenatal embryos)
Target
organ
Stage 2
Stage 3
Stage 4
Stage 5
Parent
muscle
Embryo
Gonad
Liver
4.38 1.4
77.59 24.0
18.02 2.4
3.38 1.1
0.59 1.7
0.66 1.7
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the target organ distribution was similar to that seen in the black
fin goodeid (Table 2).
3.3. VTG detection by immunohistochemistry
The ovary of the black fin goodeid is composed of an
external serous wall of connective tissue and an internal
smooth muscle layer that is supplied by capillaries, as well as
connective tissue that supports the simple cuboidal epithelial
tissue that forms long and thin folds covering most of the
ovary lumen. Pigmented cells and blood vessels are observed
in the connective tissue. The cavity of the ovary is divided by
an ovarian septum that is covered by simple cuboidal
epithelium to which is attached connective tissue with
capillaries and smooth muscle fibers. Oocytes at different
developmental stages surrounded by follicular cells forming
the granulosa and theca interna layers are located towards the
ends of the septum between the connective tissue and the
folds. After fertilization, intrafollicular gestation begins and
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Fig. 3. VTG detection in histological sections by immunohistochemistry during embryonic development of Girardinichthys viviparus. (a) External serous wall, stage
2, control (40) (b) External serous wall, stage 2, VTG-positive (40). (c) Ovarian stroma, stage 2, VTG-positive (40). (d) Spongy ovarian stroma, stage 2, VTGpositive (100). (e) Vitellogenic oocytes (stage 2), control (40). (f) Vitellogenic oocytes (stage 2), VTG-positive (40). (g) Ovarian septum, stage 3, VTG-positive
(100). (h) Trophotaeniae insertion and embryonic muscle layers, stage 3, VTG-positive (40). (i) Transverse and longitudinal sections of trophotaeniae, stage 3,
VTG-positive (40). Arrows indicate the presence and location of VTG.
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Fig. 4. VTG detection in histological sections by immunohistochemistry during embryonic development of Girardinichthys viviparus. (a) Trophotaeniae, transversal
section, stage 3, control (100). (b) Trophotaeniae, transversal section, stage 3, VTG-positive (100). (c) Panoramic of fin folding and development of dermoplacenta
with the inner wall of the ovisac, stage 4 VTG-positive (10). (d) Panoramic of the dermoplacenta of the embryo's body with ovarian stroma, stage 4 (10). (e) Fin
folding, stage 4, VTG-positive and dermoplacenta (40). (f) Body of the embryo and dermoplacenta, stage 4 control. (g) Finfold, stage 5, control (40). (h) Finfold,
stage 5, VTG-positive. (i) Head of the embryo prior to birth, VTG-positive (placentotrophy) (10). Arrows indicate the presence and location of VTG. White squares
indicate the dermoplacenta.
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