Zn(2+) induces hyperpolarization by activation of a K(+) channel and increases intracellular Ca(2+) and pH in sea urchin spermatozoa.

Zinc (Zn(2+)) has been recently recognized as a crucial element for male gamete function in many species although its detailed mechanism of action is poorly understood. In sea urchin spermatozoa, Zn(2+) was reported as an essential trace ion for efficient sperm motility initiation and the acrosome reaction by modulating intracellular pH (pHi). In this study we found that submicromolar concentrations of free Zn(2+) change membrane potential (Em) and increase the concentration of intracellular Ca(2+) ([Ca(2+)]i) and cAMP in Lytechinus pictus sperm. Our results indicate that the Zn(2+) response in sperm of this species mainly involves an Em hyperpolarization caused by K(+) channel activation. The pharmacological profile of the Zn(2+)-induced hyperpolarization indicates that the cGMP-gated K(+) selective channel (tetraKCNG/CNGK), which is crucial for speract signaling, is likely a main target for Zn(2+). Considering that Zn(2+) also induces [Ca(2+)]i fluctuations, our observations suggest that Zn(2+) activates the signaling cascade of speract, except for an increase in cGMP, and facilitates sperm motility initiation upon spawning. These findings provide new insights about the role of Zn(2+) in male gamete function.

Seasonal spermatogenesis in the Mexican endemic oviparous lizard, Sceloporus aeneus (Squamata: Phrynosomatidae).

Oviparous species of Sceloporus exhibit either seasonal or continuous spermatogenesis and populations from high-elevation show a seasonal pattern known as spring reproductive activity. We studied the spermatogenic cycle of a high-elevation (2700 m) population of endemic oviparous lizard, Sceloporus aeneus, that resided south of México, D.F. Histological analyses were performed on the testes and reproductive ducts from individual lizards collected monthly. This population of S. aeneus showed a seasonal pattern of spermatogenesis, with 4 successive phases common in other lizards. These include: 1) Quiescence in August, which contained solely spermatogonia and Sertoli cells; 2) Testicular recrudescence (September-January) when testes became active with mitotic spermatogonia, spermatocytes beginning meiosis, and the early stages of spermiogenesis with spermatids; 3) Maximum testicular activity occurred from March to May and is when the largest spermiation events ensued within the germinal epithelia, which were also dominated by spermatids and spermiogenic cells; 4) Testicular regression in June was marked with the number of all germs cells decreasing rapidly and spermatogonia dominated the seminiferous epithelium. February was a transitional month between recrudescence and maximum activity. The highest sperm abundance in the lumina of epididymides was during maximum testicular activity (March-May). Thus, before and after these months fewer spermatozoa were detected within the excurrent ducts as the testis transitions from recrudescence to maximum activity in February and from maximum activity to quiescence in June. Maximum spermatogenic activity corresponds with warmest temperatures at this study site. This pattern known as spring reproductive activity with a fall recrudescence was similar to other oviparous species of genus Sceloporus.

The ultrastructure of spermatid development during spermiogenesis within the rosebelly lizard, Sceloporus variabilis (Reptilia, Squamata, Phrynosomatidae).

Several recent studies have mapped out the characters of spermiogenesis within several species of squamates. Many of these data have shown both conserved and possibly apomorphic morphological traits that could be important in future phylogenetic analysis within Reptilia. There, however, has not been a recent study that compares spermiogenesis and its similarities or differences between two species of reptile that reside in the same genus. Thus, the present analysis details the changes to spermiogenesis in Sceloporus variabilis and then compares spermatid morphologies to that of Sceloporus bicanthalis. Many of the morphological changes that the spermatids undergo in these two species are similar or conserved, which is similar to what has been reported in other squamates. There are six main character differences that can be observed during the development of the spermatids between these two sceloporid lizards. They include the presence (S. variabilis) or absence (S. bicanthalis) of a mitochondrial/endoplasmic reticulum complex near the Golgi apparatus during acrosome development, a shallow (S. variabilis) or deep (S. bicanthalis) nuclear indentation that accommodates the acrosomal vesicle, filamentous (S. variabilis) or granular (S. bicanthalis) chromatin condensation, no spiraling (S. variabilis) or spiraling (S. bicanthalis) of chromatin during condensation, absence (S. variabilis) or presence (S. bicanthalis) of the longitudinal manchette microtubules, and the lack of (S. variabilis) or presence (S. bicanthalis) of nuclear lacunae. This is the first study that compares spermiogenic ultrastructural characters between species within the same genus. The significance of the six character differences between two distantly related species within Sceloporus is still unknown, but these data do suggest that spermiogenesis might be a good model to study the hypothesis that spermatid ontogeny is species specific.

Spermiogenesis in the imbricate alligator lizard, Barisia imbricata (Reptilia, Squamata, Anguidae).

Although the events of spermiogenesis are commonly studied in amniotes, the amount of research available for Squamata is lacking. Many studies have described the morphological characteristics of mature spermatozoa in squamates, but few detail the ultrastructural changes that occur during spermiogenesis. This study's purpose is to gain a better understanding of the subcellular events of spermatid development within the Imbricate Alligator Lizard, Barisia imbricata. The morphological data presented here represent the first complete ultrastructural study of spermiogenesis within the family Anguidae. Samples of testes from four specimens collected on the northwest side of the Nevado de Toluca, México, were prepared using standard techniques for transmission electron microscopy. Many of the ultrastructural changes occurring during spermiogenesis within B. imbricata are similar to that of other squamates (i.e., early acrosome formation, chromatin condensation, flagella formation, annulus present, and a prominent manchette). However, there are a few unique characteristics within B. imbricata spermatids that to date have not been described during spermiogenesis in other squamates. For example, penetration of the acrosomal granule into the subacrosomal space to form the basal plate of the perforatorium during round spermatid development, the clover-shaped morphology of the developing nuclear fossa of the flagellum, and the bulbous shape to the perforatorium are all unique to the Imbricate Alligator Lizard. These anatomical character differences may be valuable nontraditional data that along with more traditional matrices (such as DNA sequences and gross morphological data) may help elucidate phylogenetic relationships, which are historically considered controversial within Squamata.

Ontogenic development of spermatids during spermiogenesis in the high altitude bunchgrass lizard (Sceloporus bicanthalis).

The body of ultrastructural data on spermatid characters during spermiogenesis continues to grow in reptiles, but is still relatively limited within the squamates. This study focuses on the ontogenic events of spermiogenesis within a viviparous and continually spermatogenic lizard, from high altitude in Mexico. Between the months of June and August, testicular tissues were collected from eight spermatogenically active bunchgrass lizards (Sceloporus bicanthalis) from Nevado de Toluca, México. The testicular tissues were processed for transmission electron microscopy and analyzed to access the ultrastructural differences between spermatid generations during spermiogenesis. Interestingly, few differences exist between S. bicanthalis spermiogenesis when compared with what has been described for other saurian squamates. Degrading and coiling membrane structures similar to myelin figures were visible within the developing acrosome that are likely remnants from Golgi body vesicles. During spermiogenesis, an electron lucent area between the subacrosomal space and the acrosomal medulla was observed, which has been observed in other squamates but not accurately described. Thus, we elect to term this region the acrosomal lucent ridge. This study furthers the existing knowledge of spermatid development in squamates, which could be useful in future work on the reproductive systems in high altitude viviparous lizard species.

Are TRP channels involved in sperm development and function?

Spermatozoa must translate information from their environment and the egg to achieve fertilization in sexually reproducing animals. These tasks require decoding a variety of signals in the form of intracellular Ca(2+) changes. As TRP channels constitute a large family of versatile multi-signal transducers, they are interesting subjects in which to explore their possible participation in sperm function. Here, we review the evidence for their presence and involvement in sperm motility, maturation, and the acrosome reaction, an exocytotic process required for sperm-egg fusion. Since store-operated Ca(2+) entry (SOCE) has been proposed to play an important role in these three functions, the main proteins responsible for this transport (STIM and ORAI) and their interaction with TRPs are also discussed. Improving our tools to solve infertility, improve animal breeding, and preserve biodiversity requires a better understanding of how Ca(2+) is regulated in spermatozoa.

Temporal germ cell development strategy during continuous spermatogenesis within the montane lizard, Sceloporus bicanthalis (Squamata; Phrynosomatidae).

Sceloporus bicanthalis is a viviparous lizard that lives at higher elevations in Mexico. Adult male S. bicanthalis were collected (n = 36) from the Nevado de Toluca, Mexico (elevation is 4200 m) during August to December, 2007 and January to July, 2008. Testes were extracted, fixed in Trumps, and dehydrated in a graded series of ethanol. Tissues were embedded, sectioned (2 µm), stained, and examined via a light microscope to determine the spermatogenic developmental strategy of S. bicanthalis. In all months examined, the testes were spermiogenically active; based on this, plus the presence of sperm in the lumina of seminiferous tubules, we inferred that S. bicanthalis had year-round or continuous spermatogenesis, unlike most reptiles that occupy a temperate or montane habitat. It was recently reported that seasonally breeding reptiles had a temporal germ cell development strategy similar to amphibians, where germ cells progress through spermatogenesis as a single population, which leads to a single spermiation event. This was much different than spatial development within the testis of other derived amniotes. We hypothesized that germ cell development was temporal in S. bicanthalis. Therefore, we wanted to determine whether reptiles that practice continuous spermatogenesis have a mammalian-like spatial germ cell development, which is different than the typical temperate reptile exhibiting a temporal development. In the present study, S. bicanthalis had a temporal development strategy, despite its continuous spermatogenic cycle, making them similar to tropical anoles.

Variación morfológica de la lagartija partenogenética Aspidoscelis rodecki (Squamata: Teiidae): implicaciones evolutivas y de conservación

Morphologic variation of the parthenogenetic lizard Aspidoscelis rodecki (Squamata: Teiidae): evolutionary and conservation implications. Post-formational divergence has been used for the recognition of new parthenogenetic species. Currently, the parthenogenetic lizard Aspidoscelis rodecki McCoy and Maslin 1962 is recognized as a single taxon that was derived from a single, parthenogenetically capable, hybrid. This lizard had been derived via hybridization between individuals of two gonochoristic species, Aspidoscelis angusticeps Cope 1878 and Aspidoscelis deppii Wiegmann 1834. The distribution of A. rodecki includes Isla Contoy and Isla Mujeres and the adjacent mainland of Quintana Roo, México. Previous studies have found post-formational divergence in genetic, chromatic and life-history characteristics among a continental population (Puerto Juárez) and an insular population (Isla Contoy). A meristic analysis was carried out to evaluate the morphological divergence among both populations of A. rodecki. We used 38 individuals from Puerto Juárez and 23 individuals from Isla Contoy. Nine meristic characters with discrimination value among species of the genus Aspidoscelis were used in both univariate (t-Student) and multivariate analyses (principal components and canonical variate analysis). According to both analyses, Puerto Juárez is meristically distinguishable from Isla Contoy. Both populations differ in five meristic characters and were a high correct classification in the canonical variate analysis: 97% of Puerto Juárez and 100% of Isla Contoy. A small sample from Isla Mujeres and a single specimen from Punta Sam (mainland) may represent different morphological groups. Due to the patterns of phenotypic variation, A. rodecki is considered as a single variable parthenogenetic species with high priority to conservation. The populations of A. rodecki have been extremely affected by the tourism developers. If the habitat of the parthenogenetic lizard (beach grasses) is allowed to stay, the expansion by the developers will not affect the survivorship of these populations. Nevertheless, the first sign of development is the total destruction of natural grasses that occurs on the beach, leaving only sand. There is a last chance to save the parthenogenetic lizard A. rodecki, but any effort will be useless without the support from the environmental authority of Mexico and cooperation from the developers. We suggest that Puerto Juárez and Isla Contoy receive separate management because they have unique portions of phenotypic variation of A. rodecki. The two lizard populations can be considered separate "Evolutionary Significant Units" (ESU). Rev. Biol. Trop. 56 (4): 1871-1881. Epub 2008 December 12.

La divergencia post-formación se ha utilizado para el reconocimiento de nuevas especies partenogenéticas. Actualmente, la lagartija partenogenética Aspidoscelis rodecki McCoy y Maslin 1962 es reconocida como una sola especie, que se originó de un híbrido partenogenético. Estudios previos han encontrado divergencia genética, en coloración y en características de historia de vida entre una población continental (Puerto Juárez) y una insular (Isla Contoy) en Quintana Roo, México. Se llevó a cabo un análisis merístico para evaluar la divergencia entre ambas poblaciones de A. rodecki. Se utilizaron 38 individuos de Puerto Juárez y 23 individuos de Isla Contoy. Se usaron nueve características merísticas y se realizaron análisis univariados (t de Student) y multivariados (análisis de componentes principales y análisis de variación canónica). De acuerdo a ambos análisis, Puerto Juárez es merísticamente distinguible de Isla Contoy. Ambas poblaciones difieren en cinco características merísticas y presentaron un alto porcentaje de clasificación en el análisis de variación canónica: 97% para Puerto Juárez y 100% para Isla Contoy. Una pequeña muestra de Isla Mujeres y un solo individuo de Punta Sam (continente) pueden representar otros grupos morfológicos. Con base en los patrones de variación fenotípica, A. rodecki es considerada como una sola especie partenogenética variable y prioritaria para la conservación. Debido a que cada población contiene una porción única de la variación de A. rodecki, Puerto Juárez e Isla Contoy merecen un manejo separado y se sugiere que cada una constituya una "Unidad Evolutiva Significativa" (ESU).

[Morphologic variation of the parthenogenetic lizard Aspidoscelis rodecki (Squamata: Teiidae): evolutionary and conservation implications]. / Variación morfológica de la lagartija partenogenética Aspidoscelis rodecki (Squamata: Teiidae): implicaciones evolutivas y de conservación.

Post-formational divergence has been used for the recognition of new parthenogenetic species. Currently, the parthenogenetic lizard Aspidoscelis rodecki McCoy and Maslin 1962 is recognized as a single taxon that was derived from a single, parthenogenetically capable, hybrid. This lizard had been derived via hybridization between individuals of two gonochoristic species, Aspidoscelis ungusticeps Cope 1878 and Aspidoscelis deppii Wiegmann 1834. The distribution of A. rodecki includes Isla Contoy and Isla Mujeres and the adjacent mainland of Quintana Roo, México. Previous studies have found post-formational divergence in genetic, chromatic and life-history characteristics among a continental population (Puerto Juárez) and an insular population (Isla Contoy). A meristic analysis was carried out to evaluate the morphological divergence among both populations of A. rodecki. We used 38 individuals from Puerto Juárez and 23 individuals from Isla Contoy. Nine meristic characters with discrimination value among species of the genus Aspidoscelis were used in both univariate (t-Student) and multivariate analyses (principal components and canonical variate analysis). According to both analyses, Puerto Juárez is meristically distinguishable from Isla Contoy. Both populations differ in five meristic characters and were a high correct classification in the canonical variate analysis: 97% of Puerto Juárez and 100% of Isla Contoy. A small sample from Isla Mujeres and a single specimen from Punta Sam (mainland) may represent different morphological groups. Due to the patterns of phenotypic variation, A. rodecki is considered as a single variable parthenogenetic species with high priority to conservation. The populations of A. rodecki have been extremely affected by the tourism developers. If the habitat of the parthenogenetic lizard (beach grasses) is allowed to stay, the expansion by the developers will not affect the survivorship of these populations. Nevertheless, the first sign of development is the total destruction of natural grasses that occurs on the beach, leaving only sand. There is a last chance to save the parthenogenetic lizard A. rodecki, but any effort will be useless without the support from the environmental authority of Mexico and cooperation from the developers. We suggest that Puerto Juárez and Isla Contoy receive separate management because they have unique portions of phenotypic variation of A. rodecki. The two lizard populations can be considered separate "Evolutionary Significant Units" (ESU).