Lectin Histochemical Study of Testicular Spermatogenic Cells in Muntjak (Muntiacus muntjak muntjak)

The objective of this study was to identify the type of specific glycoconjugates and its distribution in testicular spermatogenic cells in muntjak (Muntiacus muntjak muntjak) based on lectins histochemistry. An adult male muntjak aged 4-5 years old in hard antler period was used in this study. Testicular tissue was fixed in Bouin solution and processed histologically. Histochemistry method was performed using six types biotinylated lectins such as peanut agglutinin (PNA), soybean agglutinin (SBA), wheat germ agglutinin (WGA), ricinus communis agglutinin (RCA), concanavalin A (Con A), and ulex europaeus agglutinin I (UEA I) with 20 µg/ml of concentration for PNA lectins and 15µg/ml for other type of lectins. The results showed that glycoconjugates were detected by all type of lectins except UEA I in testicular spermatogenic cells with variation in distribution pattern and also the intensity of lectins binding. Glycoconjugates β-galactose, β-glucose, mannose, Nacetylgalactosamine, N-acetylglucosamine and sialic acid were stained intensely by lectins in golgy-cap phase and acrosomal phase of spermatids. Glycoconjugate N-acetylgalactosamine was the sugar residues which distributed abundantly that marked by positive reaction with PNA, SBA, and RCA lectins. In conclusion, glycoconjugates are detected in testicular spermatids cells of muntjak indicated that glycoconjugates have an important role in spermatogenesis particularly in spermiogenesis. Key words: glycoconjugates, lectins, spermatid, spermatozoa, muntjak

STUDI HISTOKIMIA LEKTIN TERHADAP JENIS DAN DISTRIBUSI GLIKOKONJUGAT ABOMASUM KERBAU RAWA (Bubalus bubalis) KALIMANTAN SELATAN

Penelitian ini bertujuan mengkaji efisiensi pencernaan kerbau rawa dengan mengidentifikasi jenis dan distribusi glikokonjugat pada daerah abomasum kerbau rawa. Enam ekor kerbau rawa jantan >2,5 tahun dan berat badan 300-400 kg digunakan dalam penelitian ini. Sampel diperoleh dari rumah potong hewan (RPH) Kabupaten Banjar, Kalimantan Selatan. Setiap bagian abomasum meliputi kardiak, fundus, dan pilorus diambil untuk pengamatan mikroskopis dengan pewarnaan hematoksilin-eosin (HE) dan alcian blue-periodic acid schiff (AB-PAS). Residu gula glikokonjugat pada abomasum dideteksi dengan pewarnaan histokimia lektin dengan menggunakan wheat germ agglutinin (WGA), ricinus communis agglutinin (RCA), concanavalin agglutinin (Con A), ulex europaeus agglutinin (UEA), dan soybean agglutinin (SBA). Hasil penelitian menunjukkan bahwa daerah kardiak mengandung glikokonjugat D manosa/D glukosa, D galaktosa, dan N asetilglukosamin. Daerah fundus mengandung D manosa/D glukosa, D galaktosa, L fukosa, N asetilglukosamin, dan N asetilgalaktosamin. Daerah pilorus mengandung glikokonjugat L fukosa dan N asetilglukosamin. Pola reaktivitas daerah kardiak, fundus, dan pilorus kerbau rawa terhadap pewarnaan histokimia lektin memiliki pola yang berbeda dengan ruminansia lain. Jenis glikokonjugat yang dimiliki oleh kerbau rawa tersebut diduga berkaitan dengan fungsi peningkatan kemampuan efisiensi pencernaan kerbau rawa. Setiap bagian abomasum kerbau rawa memiliki jenis glikokonjugat yang spesifik dengan pola distribusi khas sesuai dengan fungsinya.

The Specific Binding between Galactose Group and Ricinus Communis Agglutinin (RCA) Investigated with Microcantilever

The specific recognization between galactose group and Ricinus Communis Agglutinin (RCA) was investigated by microcantilever. The gold side of the microcantilever was covalently bound with N-galactose, RCA and asialofetuin (ASF) via mixed self assembly monolayer of 11-mercaptoundecanoic acid and 6-mercaptohexanol, respectively. After adding RCA into the flowing cell, the deflection could be observed on the N-galactose or ASF modified microcantilever. Meanwhile, the deflection could also be observed after ASF bound to the RCA modified microcantilever. In order to prove that the deflection is caused by the specific interaction between the galactose group and RCA, bovine serum albumin (BSA) was introduced into the flowing cell as control experiment and no obvious deflection was observed. The specific interaction was also confirmed by the evidence that the bound protein layer can be mechanically removed with atomic force microscopy nanolithography technology.