The role of skeletal muscle Akt in the regulation of muscle mass and glucose homeostasis.

OBJECTIVE: Skeletal muscle insulin signaling is a major determinant of muscle growth and glucose homeostasis. Protein kinase B/Akt plays a prominent role in mediating many of the metabolic effects of insulin. Mice and humans harboring systemic loss-of-function mutations in Akt2, the most abundant Akt isoform in metabolic tissues, are glucose intolerant and insulin resistant. Since the skeletal muscle accounts for a significant amount of postprandial glucose disposal, a popular hypothesis in the diabetes field suggests that a reduction in Akt, specifically in skeletal muscle, leads to systemic glucose intolerance and insulin resistance. Despite this common belief, the specific role of skeletal muscle Akt in muscle growth and insulin sensitivity remains undefined. METHODS: We generated multiple mouse models of skeletal muscle Akt deficiency to evaluate the role of muscle Akt signaling in vivo. The effects of these genetic perturbations on muscle mass, glucose homeostasis and insulin sensitivity were assessed using both in vivo and ex vivo assays. RESULTS: Surprisingly, mice lacking Akt2 alone in skeletal muscle displayed normal skeletal muscle insulin signaling, glucose tolerance, and insulin sensitivity despite a dramatic reduction in phosphorylated Akt. In contrast, deletion of both Akt isoforms (M-AktDKO) prevented downstream signaling and resulted in muscle atrophy. Despite the absence of Akt signaling, in vivo and ex vivo insulin-stimulated glucose uptake were normal in M-AktDKO mice. Similar effects on insulin sensitivity were observed in mice with prolonged deletion (4 weeks) of both skeletal muscle Akt isoforms selectively in adulthood. Conversely, short term deletion (2 weeks) of skeletal muscle specific Akt in adult muscles impaired insulin tolerance paralleling the effect observed by acute pharmacological inhibition of Akt in vitro. Mechanistically, chronic ablation of Akt induced mitochondrial dysfunction and activation of AMPK, which was required for insulin-stimulated glucose uptake in the absence of Akt. CONCLUSIONS: Together, these data indicate that chronic reduction in Akt activity alone in skeletal muscle is not sufficient to induce insulin resistance or prevent glucose uptake in all conditions. Therefore, since insulin-stimulated glucose disposal in skeletal muscle is markedly impaired in insulin-resistant states, we hypothesize that alterations in signaling molecules in addition to skeletal muscle Akt are necessary to perturb glucose tolerance and insulin sensitivity in vivo.

Immunohistochemical detection of Tritrichomonas foetus in formalin-fixed, paraffin-embedded sections of bovine placenta and fetal lung.

An immunohistochemical technique using a monoclonal antibody was evaluated as a diagnostic tool to specifically label Tritrichomonas foetus in formalin-fixed, paraffin-embedded sections of placenta and fetal lung from bovine abortions. Trichomonads were demonstrated in tissues from each of 12 abortions due to T. foetus and none of 15 abortions due to other or unidentified causes. Moderate to marked background staining occurred only in severely autolyzed tissues from T. foetus-infected fetuses. The antibody faintly labeled 1 of 3 other species of trichomonads (Trichomonas gallinae) but did not label other protozoa, bacteria, or fungi tested.

Abortion caused by Brucella abortus biovar 1 in a free-ranging bison (Bison bison) from Yellowstone National Park.

A near-term aborted bison (Bison bison) fetus was collected near Old Faithful geyser in Yellowstone National Park, Wyoming (USA). On necropsy, the fetus liver had a small capsular tear, and there was a small quantity of blood in the peritoneal cavity. Microscopic lesions included mild, purulent bronchopneumonia and mild, multifocal, interstitial pneumonia. Brucella abortus biovar 1 was isolated from fetal abomasal contents, lung, and heart blood.

Mycobacterium bovis infection in North American elk (Cervus elaphus).

A naturally occurring outbreak of Mycobacterium bovis infection in captive wild elk (wapiti) in Montana was confirmed by mycobacteriologic examination. Twenty-eight of 143 elk responded to M. bovis purified protein derivative (PPD) tuberculin injected intradermally in the cervical region (SCT). The results of comparative cervical tuberculin skin tests conducted within 9 days of SCT revealed greater responses to M. bovis PPD tuberculin than to M. avium PPD tuberculin in 23 of 28 elk responding. At necropsy, several grossly visible tuberculous lesions were observed in the parenchyma of the lung, thoracic lymph nodes, and submandibular lymph nodes. Microscopic examination of appropriately stained tissue sections revealed the presence of granulomatous lesions containing acid-fast bacilli. An enzyme-linked immunosorbent assay (ELISA) was developed using a sarkosyl extract of M. bovis (antigen) and peroxidase-labeled protein G (conjugate); reactions were detected in the sera of 8 of 9 elk responding to M. bovis PPD tuberculin. Lymphocyte blastogenic assay responses were detected using M. bovis antigens in 7 of 9 elk positive on skin tests using M. bovis PPD.

Chlamydial-caused infectious keratoconjunctivitis in bighorn sheep of Yellowstone National Park.

An epizootic of infectious keratoconjunctivitis occurred in bighorn sheep (Ovis canadensis) in Yellowstone National Park during the winter of 1981-82. The causative organism was identified as Chlamydia sp. Mortality related to the epizootic was approximately 60% of an estimated 500 bighorn sheep in the northern range population. The infection probably affected all sex and age classes, but field surveys of live animals and mortality suggested that mature rams died disproportionately. Limited field observations the following winter on individuals having both normal and cloudy-appearing eyes suggested that half of the bighorns then present on the core units of winter range had contracted the disease and survived. By 1988, there were about 300 bighorn sheep in the population.

Fetal and placental lesions in bovine abortion due to Tritrichomonas foetus.

Laboratory records and histologic material from 13 cases of naturally occurring, early- to late-term bovine abortion due to Tritrichomonas foetus were examined. Placentitis was present in all placentas. Pyogranulomatous bronchopneumonia was in six of the 11 lungs which were examined. Trichomonad organisms were demonstrated within the placental stroma in all cases and free or phagocytized in pulmonary airways in seven cases.

Fatal neonatal toxoplasmosis in a bobcat (Lynx rufus).

Toxoplasma gondii was found in tissues of a captive 1-week-old bobcat (Lynx rufus) that died of myocarditis, hepatitis and encephalitis. Although infection is common in wild Felidae, clinical toxoplasmosis is rarely seen. In this case, the infection was apparently congenitally acquired.