Analisis isoenzim untuk mempelajari variasi genetik sapi Bali di Provinsi Bali

The aim of this research was to know Bali cattle genetic variation according to the band pattern of isoenzyme. Esterase and Malate dehydrogenase Isoenzymes of Bali cattle were studied. Using the native-PAGE method, the analysis has been made of the genetic structure and variation of the bali cattle population. Isoenzyme isolated from leucocytes cell using homogenation method by adding Phosphat Buffer Saline (PBS). A hundred sample of Bali cattles were taken in Mambang, Slemadeg and Kuwumkeladi. The result of this research indicate that from the 2 different enzyme, 3 loci were detected, and 1 of them was polymorphic (MDH-2). There was null allele phenomenom in MDH-2 locus. The loci polymorphic proportion of three population were 0,333. Chi-Square analysis of three population were 1.251–1.560. The heterozygosity value of three population (Mambang, Slemadeg and Kuwumkeladi) were 0.098, 0.111 and 0.118, respectively

Native human zona pellucida glycoproteins: Purification and binding properties

BACKGROUND: Fertilization starts with the binding of the spermatozoa to the zona pellucida (ZP) of the oocyte. Such binding is a carbohydrate-mediated event and consists of a series of tightly regulated events. Molecular interactions between spermatozoon and ZP in human are not well characterized due to limited availability of oocytes for research. Our current technology cannot generate recombinant human ZP (hZP) glycoproteins with native glycosylation. METHODS AND RESULTS: In this study, hZP glycoproteins, hZP2 (∼120 kDa), hZP3 (∼58 kDa) and hZP4 (∼65 kDa) were purified from ZP (purity >88%) by immunoaffinity columns. The binding sites of the purified native hZP3 and hZP4 were localized to the acrosome region of the capacitated human spermatozoa, and were lost after acrosome reaction. Purified human hZP2 bound to this region only in acrosome-reacted spermatozoa. Differential binding of the three glycoproteins to the post-acrosomal region and the midpiece of the spermatozoa was observed. In addition, hZP3, but not hZP2 and hZP4, induced hyperactivation. The stimulatory activity was dependent partly on N-linked glycosylation of hZP3. CONCLUSIONS: This manuscript describes the biological activities of purified hZP glycoproteins from the native source for the first time. © The Author 2008. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved.link_to_OA_fulltex

Purification of native human zona pellucida glycoproteins from eggs: Binding characteristics to human spermatozoa, hyper-activation, acrosomal exocytosis, and sperm-oocyte interaction

Abstract no. 74Fertilization starts with the binding of the spermatozoa to the zona pellucida (ZP) of the oocyte. Such binding is a carbohydrate-mediated event and consists of a series of tightly regulated events. Molecular interactions between spermatozoon and ZP in human are not well characterized due to limited availability of oocytes for research. Our current technology cannot generate recombinant human ZP glycoproteins with native glycosylation. In this study, human ZP glycoproteins, hZP2 (~120 kDa), hZP3 (~58 kDa) and hZP4 (~ 65 kDa) were purified from zonae pellucidae (purity .88%) by immunoaffinity columns. The binding sites of the purified native hZP3 and hZP4 were localized to the acrosome region of the capacitated human spermatozoa, and were lost after acrosome reaction. Purified hZP2 bound to this region only in acrosomereacted spermatozoa. Differential binding of the three glycoproteins to the post-acrosomal region and the mid-piece of the spermatozoa was observed. Both hZP3 and hZP4 induced acrosome reaction and inhibited spermatozoa-ZP binding time- and concentration-dependently to different extents. In addition, hZP3, but not hZP2 and hZP4, induced hyperactivation. These biological activities of the glycoproteins were dependent partly on their glycosylation. Both the N-linked and O-linked glycosylation contributed to the observed activities of the ZP glycoproteins in humans, though the former seemed to have a greater impact. The results describe for the first time, the biological activities of the purified human ZP glycoproteins from the native source