Takafumi Gotoh

Cellular conditions for intramuscular fat deposition in Japanese Black and Holstein steers

The experiment was conducted to study the development of intramuscular fat in Japanese Black (JB) compared to Holstein (HS) steers and to find breed differences for fat depot development and distribution in the carcass under equal feeding conditions. Additional to slaughter samples, biopsy samples of longissimus muscle (LM) and subcutaneous fat, taken at 10, 14, 18, and 22 months of age, were used for histological and molecular investigations. Japanese Black steers stored about 14% more fat in the LM (P = 0.001), resulting in larger marbling flecks (P < 0.001). Muscle fibers and intramuscular adipocytes in both breeds responded to the high energy feeding with significant enlargement, which was faster in JB. Histograms of intramuscular adipocytes size showed a shift toward larger cells during growth, but also the abundance of small, developing adipocytes. This development was accompanied by a correlated up-regulation of adipogenic genes until 22 months of age.

Different growth rates of male chicken skeletal muscles related to their histochemical properties

1. Early, M. pubo-ischio-femoralis pars medialis (PIF muscle) and late, M. iliotibialis lateralis pars postacetabularis (ITL muscle), maturing muscles were studied. These two muscles contained different populations of histochemical fibre types. 2. The profile measurements of the muscles showed diphasic allometric growth relative to the weights. In the early stage of growth (up to 15 weeks after hatching), the muscle length, width and depth all increased, while in the later stage (from 15 to 35 weeks) muscle growth resulted mainly from the marked increase in the depth and to a lesser degree, from an increase in the width. 3. Type I fibres, observed predominantly in PIF muscle matured earlier than the Type II fibres of ITL muscle. 4. From these results, the detailed process of skeletal muscle growth in the chicken was discussed.

Myofibre composition and total collagen content in M. iliotibialis lateralis and M. pectoralis of Silkie and White Leghorn chickens.

1. Using adult Silkie and White Leghorn (WL) chickens, fibre composition and total collagen content were compared between M. iliotibialis lateralis (ITL) and M. pectoralis (PT). 2. Fibres were divided into type IIR and IIW showing strong and weak reduced nicotinamide adenine dinucleotide dehydrogenase activities, respectively. 3. Fibre composition differed markedly between ITL and PT muscle. ITL muscle was composed of both types IIR and IIW fibre but PT muscle of only type IIW. Breed difference on composition was noted only in ITL muscle of cocks (type IIR; Silkie 53.8% and WL 37.0%). 4. Total collagen content was greater in ITL muscles and in cocks in both muscles. However, a breed difference was observed only in the PT muscle of hens. 5. In conclusion, the total collagen content was affected by muscle location within the body and by sex rather than by fibre composition.

Testis developments and puberty in the male Tokara (Japanese native) goat

In male Tokara (Japanese native) goats, testis development and the onset of puberty were studied histologically and observing sexual behaviour. Testes weight increased from 36+/-5.4 (S.E.) g (n=5) at 3 months of age to 126+/-6.3 g (n=6) at 12 months. The degree of the testis development differed among kids at 3 months of age and only one of four had produced spermatozoa in the testis. Large number of spermatozoa were always present in seminiferous tubules and epididymal ducts from 4 months of age. The diameter of seminiferous tubules increased from 133+/-9.9 microm (n=4) at 3 months to 198+/-1.0 microm (n=3) at 6 months with little increase thereafter. Mounting and pelvic thrusts onto a teaser doe started at from 9 to 14 weeks of age. Ejaculated semen contained sperm with good motility for the first time from 17 weeks. The male Tokara goat reaches puberty at around 4 months of age but testis development continues to 12 months.

Scanning electron microscopic observation of the architecture of collagen fibres in chicken M. iliotibialis lateralis

Abstract 1. The collagen architecture of M. iliotibialis lateralis in chicken was observed under the scanning electron microscope after muscle maceration in NaOH. 2. Immunohistochemical methods showed Type I and III collagens to be distributed over both perimysium and endomysium. 3. Thick perimysium around secondary myofibre fasciculi was composed of many large longitudinal collagen bundles and a few small circumferential bundles. In contrast, thin perimysium around primary myofibre fasciculi showed mainly circumferential bundles. 4. Endomysium had a honeycomb-like structure and consisted of a fine collagen mesh, its main fibre striation being circumferential. 5. It is suggested that functional demand differs between thick perimysium and thin endomysium.

Heterogeneous composition of histochemical fibre types in the different parts of M. longissimus thoracis from Mishima (Japanese native) steers.

In Mishima (Japanese native) steers, histochemical properties of m. longissimus thoracis were examined at 3 parts: on the level of 6th thoracic (LT I), 11th thoracic (LT II) and 5th lumbar vertebra (LT III). Myofibres were categorized into Type I, II A and II B. The same fibre type composition (I, 37%, II A, 17%, II B, 46%) was observed at LT I and LT III and another (I, 26%, II A, 15%, II B, 59%) at LT II. At each part apparent regional differences of fibre type composition could not be demonstrated except for Type I between medial and lateral subpart at LT I and Type II A between central and lateral at LT III because of its marked variation among the individual steers. Relative fibre diameter of Type I to Type II A in the combined data was large at LT I and LT III. Type II B fibres showed larger diameter than Type I only at LT III. From these results it was suggested that Mishima steers have maintained another histochemical property of the longissimus muscle differing from that of Japanese Black steers (Gotoh, Iwamoto, Ono, Nishimura, Matsuo, Nakanishi, Umetsu & Takahara, (1994). Comparative study on the regional composition of fiber types in M. Longissimus thoracis with different marbling scores for Japanese Black steers. Animal Science and Technology, 65, 454-463).

Technical note: Determination of cell-specific gene expression in bovine skeletal muscle tissue using laser microdissection and reverse-transcription quantitative polymerase chain reaction1

Skeletal muscle is a very heterogeneous tissue consisting of diverse cell types with specific transcription profiles. Therefore, the measured mRNA abundance of a certain cell type marker is influenced by the transcriptional activity as well as by the usually unknown number of contributing cells in the sample. In studies on the transcriptional activity of adipogenic genes, as indicators for the development of intramuscular adipocytes, an altered number of adipocytes or respective progenitor cells can mask changes in transcriptional activity. To overcome this problem, we started to use laser microdissection to isolate RNA of adipocytes and muscle fibers separately for downstream analysis. Even muscle fiber types can be collected and analyzed separately. Laser microdissection in combination with biopsy techniques enables gene expression studies of particular cell types during the life cycle of an animal. First experiences using laser microdissection for adipogenic gene expression studies in bovine skeletal muscle are described, and the influence of sample preparation and future challenges are discussed.

Fatty acid profiles and adipogenic gene expression of various fat depots in Japanese Black and Holstein steers

Objective of the study was to assess the breed effect on fatty acid (FA) composition of different adipose tissues and on mRNA expression of genes involved in adipogenesis and fat metabolism. Japanese Black (JB) and Holstein (HS) steers were kept under equivalent conditions with high energy intake resulting in large differences in intramuscular fat (IMF) accumulation in longissimus muscle (LM). The relative FA composition of muscle, intermuscular fat, visceral fat, and perirenal fat was comparable between JB and HS steers. Circulating fatty acids were also similar in both breeds. Most relevant breed effects were identified in IMF, underlining the uniqueness of this adipose tissue site. JB steers had more monounsaturated FA and less saturated FA. Perilipin 1 and adipose differentiation-related protein (ADFP) mRNA levels were higher in IMF of JB. The results suggest advanced maturity of IMF cells in JB and altered local conditions in muscle influencing IMF accumulation and composition.

Physiological Roles of Adipokines, Hepatokines, and Myokines in Ruminants.

Since the discovery of leptin secreted from adipocytes, specialized tissues and cells have been found that secrete the several peptides (or cytokines) that are characterized to negatively and positively regulate the metabolic process. Different types of adipokines, hepatokines, and myokines, which act as cytokines, are secreted from adipose, liver, and muscle tissue, respectively, and have been identified and examined for their physiological roles in humans and disease in animal models. Recently, various studies of these cytokines have been conducted in ruminants, including dairy cattle, beef cattle, sheep, and goat. Interestingly, a few cytokines from these tissues in ruminants play an important role in the post-parturition, lactation, and fattening (marbling) periods. Thus, understanding these hormones is important for improving nutritional management in dairy cows and beef cattle. However, to our knowledge, there have been no reviews of the characteristics of these cytokines in beef and dairy products in ruminants. In particular, lipid and glucose metabolism in adipose tissue, liver tissue, and muscle tissue are very important for energy storage, production, and synthesis, which are regulated by these cytokines in ruminant production. In this review, we summarize the physiological roles of adipokines, hepatokines, and myokines in ruminants. This discussion provides a foundation for understanding the role of cytokines in animal production of ruminants.

Localization of Tektin 1 at Both Acrosome and Flagella of Mouse and Bull Spermatozoa

Tektins (TEKTs) are constitutive filamentous proteins of microtubules in cilia, flagella, basal bodies, and centrioles. In mammals, five TEKTs (TEKT1, 2, 3, 4, and 5) have been identified in testis and spermatozoa. With the exception of TEKT1, these TEKTs have been reported to be present in spermatozoa with predominant localization at the peri-axoneme structures of flagella, i.e., mitochondria and outer dense fibers. In the present study, we produced an antibody against TEKT1 to examine the localization of TEKT1 in mouse, bull, and rat spermatozoa. By immunoblot analyses and immunofluorescence microscopy, we found TEKT1 to be present in sperm flagella and at the apical region of acrosome cap in spermatozoa of all these species. Acrosome-associated TEKT1 disappeared after in vitro acrosome reaction in mouse spermatozoa. These observations suggest another potential role for TEKT1 as a cytoskeletal element in the sperm head, or as a molecule involved in acrosome-related phenomena, such as acrosome reaction.