ABSTRACT
Cortical development requires the precise timing of neural precursor cell (NPC) terminal mitosis. Although cell cycle proteins regulate terminal mitosis, the factors that influence the cell cycle machinery are incompletely understood. Here we show in mice that myeloid cell leukemia 1 (Mcl1), an anti-apoptotic Bcl-2 protein required for the survival of NPCs, also regulates their terminal differentiation through the cell cycle regulator p27(Kip1). A BrdU-Ki67 cell profiling assay revealed that in utero electroporation of Mcl1 into NPCs in the embryonic neocortex increased NPC cell cycle exit (the leaving fraction). This was further supported by a decrease in proliferating NPCs (Pax6(+) radial glial cells and Tbr2(+) neural progenitors) and an increase in differentiating cells (Dcx(+) neuroblasts and Tbr1(+) neurons). Similarly, BrdU birth dating demonstrated that Mcl1 promotes premature NPC terminal mitosis giving rise to neurons of the deeper cortical layers, confirming their earlier birthdate. Changes in Mcl1 expression within NPCs caused concomitant changes in the levels of p27(Kip1) protein, a key regulator of NPC differentiation. Furthermore, in the absence of p27(Kip1), Mcl1 failed to induce NPC cell cycle exit, demonstrating that p27(Kip1) is required for Mcl1-mediated NPC terminal mitosis. In summary, we have identified a novel physiological role for anti-apoptotic Mcl1 in regulating NPC terminal differentiation.
Subject(s)
Brain/embryology , Brain/metabolism , Cyclin-Dependent Kinase Inhibitor p27/metabolism , Neural Stem Cells/metabolism , Proto-Oncogene Proteins c-bcl-2/metabolism , Animals , Brain/cytology , Cell Cycle Checkpoints , Cell Differentiation , Cell Proliferation , Cyclin-Dependent Kinase Inhibitor p27/deficiency , Cyclin-Dependent Kinase Inhibitor p27/genetics , Doublecortin Protein , Female , Mice , Mice, Inbred C57BL , Mice, Knockout , Mice, Transgenic , Mitosis , Myeloid Cell Leukemia Sequence 1 Protein , Neural Stem Cells/cytology , Neurogenesis , Pregnancy , Proto-Oncogene Proteins c-bcl-2/deficiency , Proto-Oncogene Proteins c-bcl-2/geneticsABSTRACT
OBJECTIVE: To report successful surgical repair of a grade IV lateral patellar luxation in a 437-kg heifer. STUDY DESIGN: Case report. ANIMAL: Seventeen-month-old Holstein heifer (437 kg). METHODS: Diagnosis of traumatic lateral patellar luxation was made based on physical examination, and confirmed on radiographs. Arthroscopic examination of the stifle assessed joint changes. Lateral patellar luxation was surgically repaired using lateral release of the patella and medial imbrication of the joint capsule. RESULTS: The heifer presented nonweight-bearing lameness of the left hind limb (5/5 lameness score). Unilateral grade IV lateral patellar luxation was diagnosed based on physical examination and radiography. Arthroscopic examination of the stifle showed synovitis and cartilage eburnation of the medial articular surface of the patella and of the lateral trochlear ridge of the femur. Lateral release of the patella and medial imbrication of the joint capsule was performed. The heifer remained lame (4.5/5 lameness score) and developed severe disuse muscle atrophy after surgery. By day 112, the heifer was walking easily and was completely weight bearing on the left hindlimb but did have a gait alteration (2/5 lameness score). On day 229, the heifer calved for the first time and lameness was no longer evident. CONCLUSION: This report documents successful surgical treatment of traumatic lateral patellar luxation in a large heifer but additional case evaluation is required to provide an accurate prognosis for this condition and treatment in large cattle.