Fluorometric Study of Rabbit Sperm Head Membrane Phospholipid Asymmetry During Capacitation and Acrosome Reaction Using Annexin-v Fitc

Ávalos-Rodrı́guez, Alejandro; Ortiz-Muñiz, Rocío; Ortega-Camarillo, Clara; Vergara‐Onofre, Marcela; Rosado-Garciá, A; Rosales‐Torres, Ana María

doi:10.1080/01485010490448741

Cited by 11 publications

(8 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It is possible that the expression level of Anex5 is higher in fESCs than in pESCs and aESCs, and that this may be related to fertilization and calcium oscillation upon sperm penetration into the oocyte. As seen in a previous report [25], after sperm capacitation, Anex5 binding sites were found mainly in the post-acrosomal region of the sperm head plasma membrane. After induction of the acrosome reaction, the Anex5 binding sites were found almost only in the acrosomal region and with higher numbers of binding sites in the equatorial area.…”

Section: Mrna and Protein Expressionsupporting

confidence: 54%

Analysis of proteomic profiling of mouse embryonic stem cells derived from fertilized, parthenogenetic and androgenetic blastocysts

Cui¹,

Shen²,

Lee³

et al. 2011

SCD

View full text Add to dashboard Cite

Embryonic stem cells (ESCs) are derived from the inner cell mass (ICM) of preimplantation embryos. ESCs exhibit true pluripotency, i.e., the ability to differentiate into cells of all three germ layers in the developing embryo. We used 2-DE MALDI-TOF/TOF to identify differentially expressed proteins among three types of mouse embryonic stem cells (ESCs) derived from fertilized, parthenogenetic, and androgenetic (fESC, pESC and aESC, respectively) blastocysts. We detected more than 800 proteins on silverstained gels of whole protein extracts from each type of ESC. Of these, 52 differentially expressed proteins were identified by the MALDI-TOF/TOF analyzer, including 32 (fESCs vs. pESCs), 28 (fESCs vs. aESCs) and 17 (pESCs vs. aESCs) prominent proteins with significantly higher or lower expression relative to the comparison cells. Among the 32 proteins from fESCs, 12 were significantly increased in expression and 20 were reduced in expression in fESCs compared with pESCs. Similarly, 10 of 28 and 8 of 17 proteins were more highly expressed in fESCs and pESCs compared with aESCs, respectively. In contrast, 18 of 28 and 9 of 17 proteins were reduced in expression in fESCs and pESCs compared with aESCs, respectively. Of the eight protein candidates in fESCs, four were increased and four were decreased in expression relative to both pESCs and aESCs in the 2-DE analysis. Differential expression of these proteins were confirmed by mRNA expression analysis using real time RT-PCR. For three proteins, ANXA5, CLIC1 and SRM, Western blot analysis corroborated the expression patterns indicated by the 2-DE results. ANXA5 and CLIC1 were increased in expression and SRM was decreased in expression in fESCs compared with both pESCs and aESCs. The differentially expressed proteins identified in the present study warrant further investigation towards the goal of their potential application in ESC therapy.

show abstract

Section: Mrna and Protein Expressionsupporting

confidence: 54%

Analysis of proteomic profiling of mouse embryonic stem cells derived from fertilized, parthenogenetic and androgenetic blastocysts

Cui¹,

Shen²,

Lee³

et al. 2011

SCD

View full text Add to dashboard Cite

show abstract

“…mexicanus spermatozoa responded satisfactorily when submitted to capacitation under the conditions utilized for other mammalian sperm cells [García-Macedo et al 2001;Á valos-Rodríguez et al 2004], i.e., incubation during 6 h at 391C under a 5% CO 2 /95% air atmosphere, in the presence of 2.5 mM Ca 2 þ . After staining with chlortetracycline, capacitated spermatozoa were easily distinguished by the presence of bright fluorescence in the acrosome region (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The acrosome reaction was induced in previously capacitated spermatozoa, either by the addition of 100 ml of A23187 diluted stock solution to a final concentration of 10 mM of the ionophore, or by the addition of progesterone to a final concentration of 3.18 mM [Sabeur et al 1996] in the capacitating medium and the incubation was continued under the same conditions. After 15, 30, 45 and 60 min at 371C, in accord with our previous results [García-Macedo et al 2001;Á valos-Rodríguez et al 2004], samples were removed, fixed, and the percentage of acrosome reacted spermatozoa was determined by CBB staining [Miller et al 1993].…”

mentioning

confidence: 99%

Spermatozoa Epididymal Maturation in the Mexican Big-Eared Bat (Corynorhinus Mexicanus)

Cervantes

Arenas‐Ríos

Ángel

et al. 2008

Systems Biology in Reproductive Medicine

View full text Add to dashboard Cite

Prolonged epididymal sperm storage in vespertilionid and rhinolophid bats, provides an interesting experimental model for the study of spermatozoa epididymal maturation. We examined the presence of the cytoplasmic droplet, and the sequential induction of capacitation and the acrosome reaction in spermatozoa obtained from different epididymal regions (caput, corpus, cauda) throughout the annual reproductive cycle of Corynorhinus mexicanus (C. mexicanus). This is a vespertilionid bat that stores spermatozoa in the epididymis for several months after testes regression. The number of sperm recovered from the different epididymal regions indicate that epididymal transit in C. mexicanus is rapid. The persistence of a high percentage of sperm cells with cytoplasmic droplet in cauda epididymis was observed in addition to a low index of capacitation and acrosome reaction in sperm cells obtained from the corpus epididymis. There was a significant increase in the percentage of capacitated and acrosome reacted spermatozoa during the storage of sperm cells in the cauda epididymis and the percentage of capacitated spermatozoa was consistently, and significantly, higher (p < 0.05) in cauda compared to the corpus epididymis at all studied dates. Tthe process of epididymal maturation in C. mexicanus is completed in the caudal region of this organ encompassing a significant period. Our results also indicate that in C. mexicanus, and in other vespertilionid and rhinolophid bats that show the same temporal asynchrony in the function of male reproductive organs, the final phases of epididymal maturation and storage are, apparently, independent of testicular function.

show abstract

“…8D and E). The tyrosine phosphorylation of sperm proteins (45,46) and the PtdSer exposure on the sperm head (47,48) that occurred during the sperm capacitation were indistinguishable between the wild-type and TMEM16E Ϫ/Ϫ sperm ( Fig. 8F and G).…”

Section: Testis Is Composed Of Several Types Of Cells (Germ Cells Sementioning

confidence: 90%

A Role of TMEM16E Carrying a Scrambling Domain in Sperm Motility

Gyobu

Miyata

Ikawa

et al. 2016

Molecular and Cellular Biology

View full text Add to dashboard Cite

cTransmembrane protein 16E (TMEM16E) belongs to the TMEM16 family of proteins that have 10 transmembrane regions and appears to localize intracellularly. Although TMEM16E mutations cause bone fragility and muscular dystrophy in humans, its biochemical function is unknown. In the TMEM16 family, TMEM16A and -16B serve as Ca 2؉ -dependent Cl ؊ channels, while TMEM16C, -16D, -16F, -16G, and -16J support Ca 2؉ -dependent phospholipid scrambling. Here, we show that TMEM16E carries a segment composed of 35 amino acids homologous to the scrambling domain in TMEM16F. When the corresponding segment of TMEM16A was replaced by this 35-amino-acid segment of TMEM16E, the chimeric molecule localized to the plasma membrane and supported Ca 2؉ -dependent scrambling. We next established TMEM16E-deficient mice, which appeared to have normal skeletal muscle. However, fertility was decreased in the males. We found that TMEM16E was expressed in germ cells in early spermatogenesis and thereafter and localized to sperm tail. TMEM16E ؊/؊ sperm showed no apparent defect in morphology, beating, mitochondrial function, capacitation, or binding to zona pellucida. However, they showed reduced motility and inefficient fertilization of cumulus-free but zona-intact eggs in vitro. Our results suggest that TMEM16E may function as a phospholipid scramblase at inner membranes and that its defect affects sperm motility.

show abstract

Fluorometric Study of Rabbit Sperm Head Membrane Phospholipid Asymmetry During Capacitation and Acrosome Reaction Using Annexin-v Fitc

Cited by 11 publications

References 26 publications

Analysis of proteomic profiling of mouse embryonic stem cells derived from fertilized, parthenogenetic and androgenetic blastocysts

Analysis of proteomic profiling of mouse embryonic stem cells derived from fertilized, parthenogenetic and androgenetic blastocysts

Spermatozoa Epididymal Maturation in the Mexican Big-Eared Bat (Corynorhinus Mexicanus)

A Role of TMEM16E Carrying a Scrambling Domain in Sperm Motility

Contact Info

Product

Resources

About