In the bone marrow, the passage of developing B cells through critical checkpoints of differentiation is associated with a reduction of specific categories of CDR3 of the Ig heavy chain (CDR‐H3), particularly those with excessive hydrophobic or charged amino acids and those with a length of eight or fewer residues. To gain insight into the role of CDR‐H3 content in the development of B cells in the spleen, we compared the sequences of V_H7183DJCμ transcripts from sorted transitional T1, marginal zone, and follicular B cell subsets to those expressed by immature IgM⁺IgD^– and mature IgM^loIgD^hi B cells in the bone marrow. Although differences in V_H utilization were noted, the T1 CDR‐H3 repertoire showed extensive similarity to that of immature bone marrow B cells, and the follicular CDR‐H3 repertoire most resembled that of mature bone marrow B cells. Unlike the splenic follicular and bone marrow mature B cell CDR‐H3 repertoires, the marginal zone B cell CDR‐H3 repertoire retained both short and highly charged amino acid motifs, including those with two arginines. Our findings suggest that antigen binding sites containing specific categories of CDR‐H3 sequence content may inhibit, permit, or even facilitate passage of the host B cell through critical checkpoints in peripheral as well as central development.